Patentiflorin A analogs as antiviral agents

ABSTRACT

The present disclosure relates to patentiflorin A analogs that are useful as antivirals, such as anti-HIV, anti-coronaviral, anti-Ebola viral, and anti-influenza viral agents and methods of use thereof.

CROSS REFERENCE OF RELATED APPLICATION

This application claims the benefit of priority of U.S. ProvisionalPatent Application Ser. No. 62/891,441 filed on Aug. 26, 2019, which ishereby incorporated by reference in its entirety for all purposes.

TECHNICAL FIELD

The present disclosure generally relates to the arylnaphthaleneglycoside derivatives, the process for their preparation, and their useas antiviral agents. More particularly, the present disclosure relatesto patentiflorin A analogs that are useful as antiviral agents, such asanti-HIV, anti-coronaviral, anti-Ebola viral, anti-Marburg viral, andanti-influenza viral agents. The present disclosure also providesmethods for treating viral infections, such as HIV, coronaviruses, Ebolavirus, Marburg virus and influenza viruses infections.

BACKGROUND

Viruses are important etiologic agents that cause infectious diseases inhumans and other mammals. They differ greatly in size, shape, chemicalcomposition, host range, and effects on hosts. After decades of studies,only a limited number of antiviral agents are available for thetreatment and/or prevention of diseases caused by viruses such as HIV,coronaviruses, Ebola, Marburg, influenza A and B and hepatitis Cviruses. Because of their toxic effects on a host, many antiviral agentsare limited in their application. Drug resistance is often very quicklydeveloped against the antiviral agent, and many viral diseases, such asHIV have no vaccines available to treat or prevent them. Accordingly,there is a need for safe and effective antiviral agents against awide-spectrum of viruses with no or low toxicity to the host.

AIDS (acquired immunodeficiency syndrome) remains one of the mostserious threats to public health. In a UNAIDS (Uniting the world againstAIDS) report, about 77 million people have been infected with the humanimmunodeficiency virus (HIV), and 37.9 million people have died fromAIDS-related illnesses since the onset of the HIV epidemic in 1981.Since the first anti-HIV drug zidovudine (AZT) was developed andapproved in 1987, more than 40 anti-HIV drugs have been formallyapproved by the U.S. Food and Drug Administration (FDA) for thetreatment of HIV infection. These drugs are categorized as nucleosidereverse transcriptase inhibitors (NRTIs), non-nucleoside reversetranscriptase inhibitors (NNRTIs), protease inhibitors (PIs), entry andfusion inhibitors, and HIV integrase strand transfer inhibitors.Although these drugs have significantly extended the lifespan ofHIV-positive people, it is worrisome that the prevalence of HIV drugresistance has increased from 11% to 29% since the global rollout of theantiretroviral therapy (ART) in 2001. In addition, the high cost andlimited availability of the current ARTs has excluded patients indeveloping countries from the benefit of combination therapies.Therefore, there is an urgent need to continuously develop novel, moreeffective, accessible, and affordable anti-HIV therapeutics.

Influenza, a severe viral infection of the respiratory system, remains amajor threat to human health. The worldwide outbreak of highlypathogenic H5N1 subtype of avian influenza virus (AIV) and the recentappearance of new type human influenza A/H1N1 have heightened publicawareness of potential global influenza pandemics. In addition todomestic poultry, AIV can also infect wild birds, pigs, cats, humans,and other animals. Three drugs, Xofluza, zanamivir and oseltamivirphosphate, have been approved for the treatment of influenza. However,the low oral bioavailability and rapid renal elimination of zanamivir,and the rapid emergence of oseltamivir-resistant influenza viruses, haveprompted the further development of more potent, longer durationtherapeutic drugs to combat potential human influenza pandemics.

Viruses belonging to Filoviridae contain minus-strand RNA as theirgenome. There are two genera, namely Marburgvirus and Ebolavirus, underthe Filoviridae family. Marburg virus is the only member in Marburgvirusgenus. There are five members in Ebolavirus genus, namely Zaireebolavirus, Sudan ebolavirus, Cote d'Ivoire ebolavirus, Restonebolavirus and Bundibugyo. Owning to their pathogenic potential, highcase mortality rate and the lack of effective therapeutics for infectedhumans, the members of family Filoviridae have been classified as“biosafety level 4” agents. The infection of filovirus may lead tohemorrhagic fever. In fact, both genera contain species that can causeepidemics of serious hemorrhagic fever in humans as well as non-humanprimates. The outbreak of Ebola virus (EBOV) disease many occurred inDemocratic Republic of the Congo. There were numerous Ebola outbreakssince 1976. The first outbreak was in 1976 at Yambuku, with 318 casesreported and 88% death rate. Later, there were two large outbreaks in1995 and 2007, in which over 250 Ebola cases were reported in eachoutbreak. In 2014-2015, West Africa experienced the largest Ebolaoutbreak. Over 28,000 cases were reported and the fatality rate reachedto 40%. Recently, an Ebola outbreak occurred again from Apr. 4, 2018. Asof May 30, 2019, a total of 1945 cases have been reported with a deathrate of 67%. There is no FDA-approved therapeutic agent specific totreat subjects infected by filovirus. The patients suffered fromfilovirus infection mainly rely on convalescent whole blood or plasmaduring Ebola outbreaks. However, this kind of empirical treatment hasmany limitations, including difficulties in mass-production as well asthe compatibility of blood group between donor and recipient. The use ofsome potential drug candidates, which include Favipiravir, ZMapp andGS-5734, are still under investigation. More clinical data is requiredto prove the safety and efficacy of these drug candidates in treatingfilovirus infection.

The emergence of novel coronavirus (SARS-CoV-2) raised internationalconcerns and scientists strive to discover potent inhibitors againstnovel coronavirus. Coronaviruses (CoVs) are enveloped, single-stranded,positive-sense RNA virus, which include Coronaviridae, Arteriviridae,and Roniviridae families. SARS-CoV-2, that causes the current COVID-19pandemic, is a β-coronavirus. There have been six CoVs identified ashuman-susceptible viruses. Two of them, SARS-CoV and MERS-CoV, couldlead to severe or even fatal respiratory tract infections. As of Aug.21, 2020, the COVID-19 epidemic has caused 797,428 deaths among over15.5 million infected cases. Several EBOV inhibitors, such asremdesivir, toremifene, and favipiravir are repurposed as anti-viralagents active against SARS-CoV-2. However, none of them have been highlyeffective to curb the COVID-19 epidemic. Highly effective viralinhibitors are thus urgently needed to combat the coronaviruses.

Natural products have been a rich source for the discovery of leadcompounds in the modern drug discovery. Justicia cf. patentiflora wasidentified as an anti-HIV plant lead through screening over 3,500 plantextracts. Bioassay-directed fractionation of the methanol extract of thestems and barks of this plant led to the isolation of three ANL(arylnaphthalene) glycoside compounds, which displayed potent inhibitoryactivity against broad HIV clinical strains with EC₅₀ values in therange of 14-37 nM [Zidovudine (AZT): 77-95 nM]. They also showedsignificant inhibitory effects against drug-resistance HIV strains.

Some arylnaphthalene lignans have been reported to have antiviralactivity in the literature. Although some of these compounds showedsignificant antiviral activities against various virus strains, theywere not considered as potential antiviral drug candidates due to theirlow selectivity indices (SIs).

There thus exists a need to develop improved antiviral agents thataddress at least some of the aforementioned needs.

SUMMARY

The present disclosure relates to a new class of patentiflorin Aanalogs, the preparation of these compounds and new intermediates, andtheir use for treatment of viral infections, such as HIV, CoV, EBOV andAIV.

In a first aspect, provided herein is a method of treating a viralinfection in a subject in need thereof, the method comprisingadministering a therapeutically effective amount of a compound to thesubject, wherein the compound has the Formula (I):

or a pharmaceutically acceptable salt or pro-drug thereof wherein,

X is oxygen or sulfur;

R¹ is R¹⁵, —OR¹⁵, —C(O)R¹⁵, or —C(O)OR¹⁵;

R², R⁵, R⁶, R¹⁰, R¹³, and R¹⁴ are each hydrogen;

R³ and R⁴ are each independently selected from the group consisting of—OR¹⁵ and —OC(O)R¹⁵; or R³ and R⁴ taken together with the carbon atomsto which they are attached form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR¹⁶;

R⁷, R⁸, and R⁹ are each independently selected from the group consistingof —OR¹⁵ and —OC(O)R¹⁵; or R⁷ and R⁸ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶; or R⁸ and R⁹ taken together with the carbon atoms towhich they are attached to form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR¹⁶;

R¹¹ and R¹² taken together form oxo; or while one of R¹¹ and R¹² ishydrogen or halogen, the other one of R¹¹ and R¹² is selected from thegroup consisting of R¹⁵, —OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹⁵ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, heteroaryl, —OR⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N═C(R⁷)R¹⁸, —N(R¹⁷)R¹⁸,—N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N(R¹⁷)S(O)₂R¹⁸, 1,3,2-dioxaborolaneoptionally substituted with 1, 2, 3, or 4 group(s) independentlyselected from alkyl, a glycosidic group, alkynyl optionally substitutedwith a trialkylsilane, hydrocarbyl optionally substituted with 1, 2, 3,4 or 5 group(s) independently selected from R¹⁶, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶, and —(CH₂)_(k)-heterocyclyl optionally substitutedwith 1, 2, 3, 4 or 5 group(s) independently selected from R¹⁶, wherein kis an integer between 1-6;

R¹⁶ for each occurrence is independently selected from the groupconsisting of alkynyl, halogen, trichloromethyl, trifluoromethyl, cyano,nitro, oxo, ═NR¹⁷, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁷, —N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸,—N(R¹⁷)C(O)R¹⁸ and —N(R⁷)S(O)₂R¹⁸; and

R¹⁷ and R¹⁸ for each occurrence are independently hydrogen, alkyl,alkynyl, cycloalkyl, aryl, or heteroaryl, or selected from hydrocarbyland heterocyclyl, either of which is optionally substituted with 1, 2,3, 4 or 5 group(s) independently selected from the group consisting ofhalogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆ alkoxy;

with the proviso that the compound of Formula (I) is not a compoundselected from the group consisting of 5, 15a, 15b, 16, 17b, 19d,25a-25g, and 26a-26g:

In certain embodiments of the method, each of R³ and R⁴ is —O-alkyl; R⁸and R⁹ taken together with the carbon atoms to which they are attachedform a methylenedioxy ring;

R¹ is heteroaryl, —OR¹⁵, —C(O)R¹⁵, —N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸,—N═C(R¹⁷)R¹⁸, pinacolboryl, —OS(O)₂CF₃, a glycosidic group,heterocyclcyl optionally substituted with 1 or 2 group(s) independentlyselected from R¹¹, or alkynyl optionally substituted with atrialkylsilane; or

R¹ is —O(CH₂)_(m)-cyano, —O(CH₂)_(m)-alkynyl, —O(CH₂)_(m)—C(O)N(R¹⁷)R¹⁸,or —O(CH₂)_(m)—C(O)OR¹⁷, wherein m is a whole number elected from 1-4;and R¹¹ and R¹² taken together form oxo.

In certain embodiments of the method, the compound is selected from thegroup consisting of 12a, 12b, 13, 14a, 14b, 17a, 17c, 17d, 17e, 17f,17g, 17h, 18, 19a, 19b, 19c, 19e, 19f, 19g, 19h, 19i, 19j, 19k, 19l,19m, 19n, 19o, 19p, 20, 21, 22, 23 and 24:

In certain embodiments of the method, the viral infection is selectedfrom the group consisting of HIV, coronaviruses, influenza viruses,Ebola virus, and Marburg virus.

In certain embodiments of the method, R¹ is alkynyl, pinacolboryl,—OCH₂-cyano, —OCH₂—C(O)N(R¹⁷)R¹⁸, or —OCH₂—C(O)OR¹⁷; each of R³ and R⁴is —OCH₃; R⁸ and R⁹ taken together with the carbon atoms to which theyare attached form a methylenedioxy ring; R¹¹ and R¹² taken together formoxo; and the viral infection is HIV.

In certain embodiments of the method, R¹ is a glycosidic grouprepresented by the Formula (V):

wherein, R¹⁹ and R²⁰ taken together to form oxo; or while one of R¹⁹ andR²⁰ is hydrogen or halogen, the other one of R¹⁹ and R²⁰ is selectedfrom the group consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷;

R²¹ and R²² taken together to form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R²¹ and R²² is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²⁰ and R²² taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR¹⁶;

R²³ and R²⁴ taken together to form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR⁶;

R²⁵ and R²⁶ taken together form oxo; or while one of R²⁵ and R²⁶ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)OR²⁷,—CH₂R²⁷, and —C(O)R²⁷; or R²⁴ and R²⁶ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R⁶;

R²⁷ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, —OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹,—S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰,—N(R²⁹)C(O)R³⁰, —N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with1, 2, 3, 4 or 5 group(s) independently selected from R²⁸, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R²⁸, and —(CH₂)_(k)-heterocyclyl optionally substitutedwith 1, 2, 3, 4 or 5 group(s) independently selected from R²⁸, wherein kis an integer between 1-6;

R²⁸ for each occurrence is independently selected from halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³¹ and—N(R²⁹)S(O)₂R³⁰; and

R²⁹ and R³⁰ for each occurrence are each independently hydrogen orselected from hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy.

In certain embodiments of the method, R¹⁹, R²¹, R²³, and R²⁵ are eachhydrogen; R²⁰, R²² and R²⁴ are each independently selected from thegroup consisting of —OR²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)R²⁷ and —OC(O)OR²⁷;and R²⁶ is hydrogen, methyl, —OR²⁷, —OC(O)R²⁷ or —CH₂—OC(O)R²⁷; or R²⁰and R²² taken together with the carbon atoms to which they are attachedform a 5-6 membered heterocyclyl, which is optionally substituted with1, 2, 3, 4 or 5 group(s) independently selected from C₁₋₆ alkyl and C₁₋₆alkoxy; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached form a 5-6 membered heterocyclyl, which is optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromC₁₋₆ alkyl and C₁₋₆ alkoxy; or R²⁴ and R²⁶ taken together with thecarbon atoms to which they are attached form a 5-6 memberedheterocyclyl, which is optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy; and theglycloside is a monosaccaride.

In certain embodiments of the method, the glycloside is a monosaccharideselected from the group consisting of an α-L isomer and an β-L isomer.

In certain embodiments of the method, the compound is selected from thegroup consisting of 27aa, 27ab, 27ac, 27ad, 27ae, 27af, 27ba, 27bb,27bc, 27bd, 27be, 27bf, 28ab1, 28ab2, 28ab3, 28bb1, 28bb2, 28bb3, 29a,29b, 30a, 30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i, 30j, 30k, 30l, 31a,31b, 31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, 31k, 31l, 32a, 32b, 32c and32d:

In certain embodiments of the method, the viral infection is selectedfrom the group consisting of HIV, coronaviruses, influenza viruses,Ebola virus, and Marburg virus.

In certain embodiments of the method, each of R³ and R⁴ is —OCH₃; R⁸ andR⁹ taken together with the carbon atoms to which they are attached forma methylenedioxy ring; R¹¹ and R¹² taken together form oxo; R¹⁹, R²¹,R²³, and R²⁵ are each hydrogen; R²⁰, R²², and R²⁴ are each independentlyselected from the group consisting of —OR²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)R²⁷and —OC(O)OR²⁷; R²⁶ is hydrogen, methyl, —OR²⁷, —OC(O)R²⁷ or—CH₂—OC(O)R²⁷; or R²⁰ and R²² taken together with the carbon atoms towhich they are attached form a 5-6 membered heterocyclyl, which isoptionally substituted with 1 or 2 group(s) independently selected fromC₁₋₆ alkyl; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached form a 5-6 membered heterocyclyl, which is optionallysubstituted with 1 or 2 group(s) independently selected from C₁₋₆ alkyl;or R²⁴ and R²⁶ taken together with the carbon atoms to which they areattached form a 5-6 membered heterocyclyl, which is optionallysubstituted with 1 or 2 group(s) independently selected from C₁₋₆ alkyl;and the glycloside is a monosaccaride; and the viral infection is HIV,coronaviruses, influenza viruses, Ebola virus, or Marburg virus.

In a second aspect, provided herein is a compound of Formula (I):

or a pharmaceutically acceptable salt or pro-drug thereof wherein,

X is oxygen or sulfur;

R¹ is R¹⁵, —OR¹⁵, —C(O)R¹⁵, or —C(O)OR¹⁵;

R², R⁵, R⁶, R¹⁰, R¹³, and R¹⁴ are each hydrogen;

R³ and R⁴ are each independently selected from the group consisting of—OR⁵ and —OC(O)R¹⁵; or R³ and R⁴ taken together with the carbon atoms towhich they are attached form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromC₁₋₆ alkyl and C₁₋₆ alkoxy;

R⁷, R⁸, and R⁹ are each independently selected from the group consistingof —OR¹⁵ and —OC(O)R¹⁵; or R⁷ and R⁸ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from C₁₋₆ alkyl and C₁₋₆ alkoxy; or R⁸ and R⁹ taken togetherwith the carbon atoms to which they are attached to form a 5-6 memberedheterocyclyl optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy;

R¹¹ and R¹² taken together form oxo; or while one of R¹¹ and R¹² ishydrogen or halogen, the other one of R¹¹ and R¹² is selected from thegroup consisting of R¹⁵, —OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹⁵ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, heteroaryl, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N═C(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸,—N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N(R¹⁷)S(O)₂R¹⁸, 1,3,2-dioxaborolaneoptionally substituted with 1, 2, 3, or 4 group(s) independentlyselected from alkyl, a glycosidic group, alkynyl optionally substitutedwith a trialkylsilane, hydrocarbyl optionally substituted with 1, 2, 3,4 or 5 group(s) independently selected from R¹⁶, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶, and —(CH₂)_(k)-heterocyclyl optionally substitutedwith 1, 2, 3, 4 or 5 group(s) independently selected from R¹⁶, wherein kis an integer between 1-6;

R¹⁶ for each occurrence is independently selected from the groupconsisting of alkynyl, halogen, trichloromethyl, trifluoromethyl, cyano,nitro, oxo, ═NR¹⁷, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸,—N(R¹⁷)C(O)R¹⁸ and —N(R¹⁷)S(O)₂R¹⁸; and

R¹⁷ and R¹⁸ for each occurrence are independently hydrogen, alkyl,alkynyl, cycloalkyl, aryl, or heteroaryl, or selected from hydrocarbyland heterocyclyl, either of which is optionally substituted with 1, 2,3, 4 or 5 group(s) independently selected from the group consisting ofhalogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆ alkoxy,

with the proviso that the compound of Formula (I) is not a compoundselected from the group consisting of 5, 15a, 15b, 16, 17b, 19d,25a-25g, and 26a-26g:

In certain embodiments, each of R³ and R⁴ is —O-alkyl; R⁸ and R⁹ takentogether with the carbon atoms to which they are attached form amethylenedioxy ring; and R¹¹ and R¹² taken together form oxo.

In certain embodiments of the compound, R¹ is heteroaryl, —OR¹⁵,—C(O)R¹⁵, —N(R⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N═C(R¹⁷)R¹⁸, pinacolboryl,—OS(O)₂CF₃, alkynyl optionally substituted with a trialkylsilane, orheterocyclcyl optionally substituted with 1 or 2 group(s) independentlyselected from R¹⁶; or R¹ is —O(CH₂)_(m)-cyano, —O(CH₂)_(m)-alkynyl,—O(CH₂)_(m)—C(O)N(R¹⁷)R¹⁸, or —O(CH₂)_(m)—C(O)OR¹⁷, wherein m is a wholenumber selected from 1-4.

In certain embodiments of the compound, R¹ is heteroaryl, —OR¹⁵,—C(O)R¹⁵, —N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N═C(R¹⁷)R¹⁸, pinacolboryl,—OS(O)₂CF₃, heterocyclcyl optionally substituted with 1 or 2 group(s)independently selected from R¹⁶, or alkynyl optionally substituted witha trialkylsilane; or R¹ is —O(CH₂)_(m)-cyano, —O(CH₂)_(m)-alkynyl,—O(CH₂)_(m)—C(O)N(R¹⁷)R¹⁸, or —O(CH₂)_(m)—C(O)OR¹⁷, wherein m is a wholenumber selected from 1-4; each of R³ and R⁴ is —OMe; R⁸ and R⁹ takentogether with the carbon atoms to which they are attached form amethylenedioxy ring; and R¹¹ and R¹² taken together form oxo.

In certain embodiments of the compound, R¹ is alkynyl, pinacolboryl;—N(R¹⁷)R¹⁸, —OCH₂-cyano, —OCH₂-alkynyl, —OCH₂—C(O)N(R¹⁷)R¹⁸, or—OCH₂—C(O)OR¹⁷.

In certain embodiments of the compound, the compound is selected fromthe group consisting of 12a, 12b, 13, 14a, 14b, 17a, 17c, 17d, 17e, 17f,17g, 17h, 18, 19a, 19b, 19c, 19e, 19f, 19g, 19h, 19i, 19j, 19k, 19l,19m, 19n, 19o, 19p, 20, 21, 22, 23 and 24:

In certain embodiments of the compound, R¹ is a glycosidic groupselected from the group consisting of a monosaccharide, disaccharide,trisaccharide, tetrasaccharide and polysaccharide group containing lessthan 30 monosaccharides, wherein the glycosidic group comprises one ormore monosaccharides isomers selected from the group consisting of α-D,α-L, β-D, and β-L.

In certain embodiments of the compound, R¹ is a glycosidic grouprepresented by the Formula (V):

wherein, R¹⁹ and R²⁰ taken together to form oxo; or while one of R¹⁹ andR²⁰ is hydrogen or halogen, the other one of R¹⁹ and R²⁰ is selectedfrom the group consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷;

R²¹ and R²² taken together to form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R²¹ and R²² is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²⁰ and R²² taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl, which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from C₁₋₆ alkyl and C₁₋₆ alkoxy;

R²³ and R²⁴ taken together to form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl, which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from C₁₋₆ alkyl and C₁₋₆ alkoxy;

R²⁵ and R²⁶ taken together to form oxo; or while one of R²⁴ and R²⁵ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)OR²⁷,—CH₂R²⁷, and —C(O)R²⁷; or R²⁴ and R²⁶ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyl,which is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy;

R²⁷ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, —OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹,—S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰,—N(R²⁹)C(O)R³⁰, —N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with1, 2, 3, 4 or 5 group(s) independently selected from R²⁸, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R²⁸, and —(CH₂)_(k)-heterocyclyl optionally substitutedwith 1, 2, 3, 4 or 5 group(s) independently selected from R²⁸, wherein kis an integer between 1-6;

R²⁸ for each occurrence is independently selected from halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰ and—N(R²⁹)S(O)₂R³⁰; and

R²⁹ and R³⁰ for each occurrence are each independently hydrogen orselected from hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy.

In certain embodiments of the compound, R¹⁹, R²¹, R²³, and R²⁵ are eachhydrogen; R²⁰, R²², and R²⁴ are each independently selected from thegroup consisting of —OR²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)R²⁷ and —OC(O)OR²⁷;and R²⁶ is hydrogen, methyl, —OR²⁷, —OC(O)R²⁷, or —CH₂—OC(O)R²⁷; or R²⁰and R²² taken together with the carbon atoms to which they are attachedform a 5-6 membered heterocyclyl, which is optionally substituted with1, 2, 3, 4 or 5 group(s) independently selected from C₁₋₆ alkyl and C₁₋₆alkoxy; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached form a 5-6 membered heterocyclyl, which is optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromC₁₋₆ alkyl and C₁₋₆ alkoxy; or R²⁴ and R²⁶ taken together with thecarbon atoms to which they are attached form a 5-6 memberedheterocyclyl, which is optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy; and theglycloside is a monosaccaride.

In certain embodiments of the compound, the glycloside is amonosaccharide selected from the group consisting of an α-L isomer andan β-L isomer.

In certain embodiments of the compound, the compound is selected fromthe group consisting of 27aa, 27ab, 27ac, 27ad, 27ae, 27af, 27ba, 27bb,27bc, 27bd, 27be, 27bf, 28ab1, 28ab2, 28ab3, 28bb1, 28bb2, 28bb3, 29a,29b, 30a, 30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i, 30j, 30k, 30l, 31a,31b, 31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, 31k, 31l, 32a, 32b, 32c and32d:

In a third aspect, provided herein is a pharmaceutical compositioncomprising a compound as described herein and at least onepharmaceutically acceptable excipient.

Also provided herein is an arylnaphthalene lignan compounds having thegeneral Formula (I) or (II) or a pharmaceutically acceptable salt orpro-drug thereof, for use in the treatment, prevention or delay ofprogression of a virus infection in a patient.

or a stereoisomer thereof, or an enantiomer thereof, or apharmaceutically acceptable salt or pro-drug thereof, wherein

X is oxygen or sulfur;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹ and R¹⁰ are each independentlyhydrogen, halogen or a moiety comprising 1 to 30 plural valence atomsselected from carbon, boron, nitrogen, oxygen, silicon and sulfur; or R²and R³, R³ and R⁴, R⁴ and R⁵, R⁶ and R⁷, R⁷ and R⁸, R⁸ and R⁹ or R⁹ andR¹⁰ may be taken together with the carbon atoms to which they areattached to form a cyclic group which is optionally substituted with 1,2, 3, 4 or 5 group(s) independently selected from halogen and a moietycomprising 1 to 30 plural valence atoms selected from carbon, boron,nitrogen, oxygen, silicon, and sulfur;

R¹¹ and R¹² taken together form oxo; or while one of R¹¹ and R¹² ishydrogen or halogen, the other one of R¹¹ and R¹² is selected from R¹⁵,—OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹³ and R¹⁴ taken together form oxo; or while one of R¹³ and R¹⁴ ishydrogen or halogen, the other one of R¹³ and R¹⁴ is selected from R¹⁵,—OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹⁵ is independently selected from hydrogen, halogen, trichloromethyl,trifluoromethyl, cyano, nitro, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸,—C(O)OR¹⁷, —OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸,—N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N(R¹⁷)S(O)₂R¹⁸, hydrocarbyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶, heterocyclcyl optionally substituted with 1, 2, 3, 4or 5 group(s) independently selected from R¹⁶, and —(CH₂)-heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶, wherein k is an integer between 1 and 6 (e.g. 1, 2 or3);

R¹⁶ is independently selected from halogen, trichloromethyl,trifluoromethyl, cyano, nitro, oxo, ═NR¹⁷, —OR¹⁷, —C(O)R¹⁸,—C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷, —OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸,—N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸ and —N(R¹⁷)S(O)₂R¹⁸;

R¹⁷ and R¹⁸ are each independently hydrogen or selected from hydrocarbyland heterocyclyl, either of which is optionally substituted with 1, 2,3, 4 or 5 group(s) independently selected from halogen, cyano, amino,hydroxy, C₁₋₆ alkyl and C₁₋₆ alkoxy; and

R is hydrogen or selected from hydrocarbyl and heterocyclyl, either ofwhich is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyland C₁₋₆ alkoxy.

The present disclosure also provides an arylnaphthalene lignan compoundhaving the general Formula (III) or (IV) or a pharmaceuticallyacceptable salt or pro-drug thereof, for use in the treatment,prevention or delay of progression of a virus infection in a patient.

or a stereoisomer thereof, or an enantiomer thereof; or apharmaceutically acceptable salt or pro-drug thereof wherein,

X is oxygen or sulfur;

the glycosidic group is generally a carbohydrate group, especially amonosaccharide, disaccharide, trisaccharide, tetrasaccharide orpolysaccharide group, and may exist in various isomeric forms, forexample α-D, α-L, β-D or β-L forms; the carbohydrate group may beoptionally substituted with other type of substituents or evenadditional glycosidic groups; the total number of monosaccharide andsubstituted monosaccharide contained in the chemical structure of thecompound of Formula (III) and Formula (IV) may not exceed 30;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸ and R⁹ are each independently hydrogen,halogen or a moiety comprising 1 to 30 plural valence atoms selectedfrom carbon, nitrogen, oxygen and sulfur; or R¹ and R², R² and R³, R³and R⁴, R⁵ and R⁶, R⁶ and R⁷, R⁷ and R⁸, R⁸ and R⁹ may be taken togetherwith the carbon atoms to which they are attached to form a cyclic groupwhich is optionally substituted with halogen or a moiety comprising 1 to30 plural valence atoms selected from carbon, nitrogen, oxygen andsulfur;

R¹⁰ and R¹¹ may be taken together to form oxo; or while one of R¹⁰ andR¹¹ is hydrogen or halogen, the other one of R¹⁰ and R¹¹ is selectedfrom the group consisting of R²⁷, —OR²⁷, —C(O)R²⁷, and —C(O)OR²⁷;

R¹² and R¹³ may be taken together to form oxo; or while one of R¹² andR¹³ is hydrogen or halogen, the other one of R¹² and R¹³ is selectedfrom the group consisting of R²⁷, —OR²⁷, —C(O)R²⁷, and —C(O)OR²⁷;

R¹⁹ and R²⁰ taken together to form oxo; or while one of R¹⁹ and R²⁰ ishydrogen or halogen, the other one of R¹⁹ and R²⁰ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²¹ and R²² taken together to form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R²¹ and R²² is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²³ and R²⁴ taken together form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²⁵ and R²⁶ taken together form oxo; or while one of R²⁵ and R²⁶ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from R²⁷,—OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, —CH₂R³¹ and —C(O)R³¹;

R²⁷ is independently selected from hydrogen, halogen, trichloromethyl,trifluoromethyl, cyano, nitro, —OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰,—C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰,—N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰, and —N(R²⁹)S(O)₂R³⁰, hydrocarbyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from the group consisting of R²⁸, heterocyclcyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromthe group consisting of R²⁸, and —(CH₂)_(k)-heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromthe group consisting of R²⁸, wherein k is an integer between 1 and 6(e.g. 1, 2 or 3);

R²⁸ is independently selected from halogen, trichloromethyl,trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹, —C(O)R³⁰,—C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰,—N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰, and —N(R²⁹)S(O)₂R³⁰;

R²⁹ and R³⁰ are each independently hydrogen or selected from hydrocarbyland heterocyclyl, either of which is optionally substituted with 1, 2,3, 4 or 5 group(s) independently selected from the group consisting ofhalogen, cyano, amino, hydroxy, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R³¹ is independently selected from hydrogen, halogen, trichloromethyl,trifluoromethyl, cyano, nitro, —OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰,—C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰,—N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰, —N(R²⁹)S(O)₂R³⁰, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, substitutedtetrasaccharide, hydrocarbyl optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from the group consisting of R²⁸, andheterocyclyl optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from the group consisting of R²⁸; and

R is hydrogen or selected from hydrocarbyl and heterocyclyl, either ofwhich is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyland C₁₋₆ alkoxy.

A fourth aspect of the invention is a pharmaceutical formulationcomprising an arylnaphthalene lignan compound, or a pharmaceuticallyacceptable salt or prodrug thereof, for use in the treatment, preventionor delay of progression of a viral infection in a patient.

A fifth aspect of the invention is a pharmaceutical formulationcomprising a glycosidic arylnaphthalene lignan compound, or apharmaceutically acceptable salt or prodrug thereof, for use in thetreatment, prevention or delay of progression of a viral infection in apatient.

A sixth aspect of the invention is a pharmaceutical formulationcomprising a diphyllin analog, or a pharmaceutically acceptable salt orprodrug thereof, for use in the treatment, prevention or delay ofprogression of a viral infection in a patient.

A seventh aspect of the invention is a pharmaceutical formulationcomprising a patentiflorin A analog, or a pharmaceutically acceptablesalt or prodrug thereof, for use in the treatment, prevention or delayof progression of a viral infection in a patient.

Another aspect of the invention concerns the method to provide synthesisof new arylnaphthalene lignan compounds as well as the intermediatecompounds during the synthesis. In addition, the invention is directedto an intermediary compound useful in preparing other compounds of theinvention.

Compounds of the invention may exist in different forms, such as freeacids, free bases, esters and other prodrugs, salts and tautomers, andthe disclosure includes all variant forms of these compounds.

The extent of protection includes counterfeit or fraudulent productswhich contain or purport to contain a compound of the inventionirrespective of whether they do in fact contain such a compound andirrespective of whether any such compound is contained in atherapeutically effective amount.

Included in the scope of protection are packages which include adescription or instructions which indicate that the package contains aspecies or pharmaceutical formulation of the invention and a productwhich is or comprises, or purports to be or comprise, such a formulationor species. Such packages may be, but are not necessarily, counterfeitor fraudulent.

Features, integers, characteristics, compounds, chemical moieties orgroups described in conjunction with a particular aspect, embodiment orexample of the invention are to be understood to be applicable to anyother aspect, embodiment or example described herein unless incompatibletherewith.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the structure and carbon numbering of patentiflorin A andschematic preparation of diphyllin (5).

FIG. 2 shows schematic preparation of diphyllin analogs 12-17 (forpreparation of compound 14a, the reaction reagents and conditions of a:NH₄Cl, HATU, Et₃N, CH₂Cl₂; for preparation of compound 14b, the reactionreagents and conditions of b: MeNH₂, EDCI, HOBt and THF; for preparationof compound 15a, the reaction reagents and conditions of c: PdCl₂,K₂CO₃, TBAB, H₂O, 60° C.; for preparation of compound 15b, the reactionreagents and conditions of d: Cs₂CO₃, DMF, 120° C.; for preparation ofcompounds 17a-17c, the reaction reagents and conditions of e: Cs₂CO₃,acetone, r.t.; for preparation of compounds 17d, the reaction reagentsand conditions of f: K₂CO₃, DMF, 100° C.; for preparation of compounds17e-17h, the reaction reagents and conditions of g: Et₃N, DMAP, CH₂Cl₂,25° C.).

FIG. 3 shows schematic preparation of diphyllin analogs 18-24 (forpreparation of compound 19a, the reaction reagents and conditions of a:[Pd(dppf)Cl₂]CH₂Cl₂, KOAc, B₂(Pin)₂, dioxane, 100° C.; for preparationof compound 19b, the reaction reagents and conditions of b:[Pd(dppf)Cl₂]CH₂Cl₂, AlMe₃, toluene, 100° C.; for preparation ofcompound 19c, the reaction reagents and conditions of c:[Pd(dppf)Cl₂]CH₂Cl₂, Na₂CO₃, pyridine-4-boronic acid, H₂O, Dioxane, 100°C.; for preparation of compound 19d, the reaction reagents andconditions of d: Pd(OAc)₂, PPh₃, Et₃N, HCO₂H, DMF, 100° C.; forpreparation of compounds 19e and 19f, the reaction reagents andconditions of e: Pd(OAc)₂, BINAP, Cs₂CO₃, RH, toluene, 100° C.; forpreparation of compound 19g, the reaction reagents and conditions of f:Pd(PPh₃)₄, Et₂Zn, THF, 100° C.; for preparation of compounds 19h-19j,the reaction reagents and conditions of g: Pd(PPh₃)₄, K₂CO₃, RB(OH)₂,DMF, 100° C.; for preparation of compound 19k, the reaction reagents andconditions of h: Pd(PPh₃)₄, Na₂CO₃, furan-2-boronic acid, H₂O, Dioxane,100° C.; for preparation of compound 19l, the reaction reagents andconditions of i: Pd(PPh₃)₄, Na₂CO₃,1-Boc-3,6-dihydro-2H-pyridine-4-boronic acid pinacol ester, H₂O,Dioxane, 100° C.; for preparation of compound 19m, the reaction reagentsand conditions of j: Pd₂(dba)₃, XantPhos, Cs₂CO₃, morpholine DMF, 100°C.; for preparation of compound 19n, the reaction reagents andconditions of k: Pd(PPh₃)₂Cl₂, tributyl(1-ethoxyvinyl)tin, PhMe, 100°C., followed by 50% HCl; for preparation of compound 19o, the reactionreagents and conditions of l: Pd(PPh₃)₂Cl₂, CuI, Et₃N, DMF,trimethylsilylacetylene, 80° C.; for preparation of compound 19p, thereaction reagents and conditions of m: Pd(OAc)₂, Cs₂CO₃, Xantphos, Et₃N,benzophenone imine, dioxane, 100° C.; for preparation of compound 20,the reaction reagents and conditions of n: NaBH₄, MeOH, rt; forpreparation of compound 21, the reaction reagents and conditions of o:K₂CO₃, MeOH, r.t.; for preparation of compound 22, the reaction reagentsand conditions of p: HCl, THF, r.t., followed by KOH, r.t.; forpreparation of compound 23, the reaction reagents and conditions of q:Cs₂CO₃, propargyl bromide, MeCN, r.t.; for preparation of compound 24,the reaction reagents and conditions of r: Ac₂O, pyridine, r.t.).

FIG. 4 shows schematic preparation of patentiflorin A analogs 25a-25gand 26a-26g. Compounds 26a, 26b, 26c, 26d, 26e, 26f and 26g are producedfrom 25a, 25b, 25c, 25d, 25e, 25f and 25g, respectively.

FIG. 5 shows schematic preparation of patentiflorin A analogs 27aa-27af,27ba-27bf, 28ab1-28ab3, 28bb1-28bb3, 29a, 29b, 30a-30l and 31a-31l (forpreparation of compounds 27aa-27af and 27ba-27bf, the reaction reagentsand conditions of a: Ac₂O, TBAOAc, MeCN, 40° C.; for preparation ofcompounds 28ab1 to 28ab3 and 28bb1 to 28bb3, the reaction reagents andconditions of b: AllocOCl, Et₃N, CH₂Cl₂, 0° C. to r.t.; for preparationof compounds 29a and 29b, the reaction reagents and conditions of c:AcCl, CH₂Cl₂, MeOH, 0° C. to r.t.; for preparation of compounds 30a-k,the reaction reagents and conditions of d: Et₃N, DMAP, dry CH₂Cl₂, 0° C.to r.t.; for preparation of compounds 30l, the reaction reagents andconditions of e: KOH, 18-Crown-6, THF, r.t.; for preparation ofcompounds 31a-1, the reaction reagents and conditions of f: (PPh₃)₄Pd,PPh₃, Et₃N, HCOOH, THF, N₂, 55° C.). Compounds 27aa, 27ab, 27ac, 27ad,27ae and 27af are produced from 26a. Compounds 27ba, 27bb, 27bc, 27bd,27be and 27bf are produced from 26b. Compounds 28ab1, 28ab2 and 28ab3are produced from 27ab. Compounds 28bb1, 28bb2 and 28bb3 are producedfrom 27bb. Compounds 29a and 29b are produced from 28ab3 and 28bb3,respectively. Compounds 30a and 30l are produced from 29a. Compounds30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i, 30j and 30k are produced from29b. Compounds 31a, 31b, 31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, 31k and31l are produced from 30a, 30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i, 30j,30k and 30l, respectively.

FIG. 6 shows schematic preparation of patentiflorin A analogs 32a-32d.Compounds 32a is produced from 26f. Compounds 32b-32d are produced from26g.

DETAILED DESCRIPTION

The present disclosure is not to be limited in scope by any of thespecific embodiments described herein. The following embodiments arepresented for exemplification only.

Throughout the description and claims of this specification the word“comprise” and other forms of the word such as “comprising” and“comprises,” means including but not limited to, and is not intended toexclude, for example, other additives, components, integers, or steps.

As used in the description and the appended claims, the singular forms“a,” “an,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “a composition”includes mixtures of two or more such compositions, reference to “thecompound” includes mixtures of two or more such compounds, reference to“an agent” includes mixture of two or more such agents, and the like.

“Optional” or “optionally” means that the subsequently described eventor circumstance can or cannot occur, and that the description includesinstances where the event or circumstance occurs and instances where itdoes not.

As used herein by a “subject” is meant an individual. Thus, the“subject” can include domesticated animals (e.g., cats, dogs, etc.),livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratoryanimals (e.g., mouse, rabbit, rat, guinea pig, etc.), and birds.“Subject” can also include a mammal, such as a primate or a human.

By “reduce” or other forms of the word, such as “reducing” or“reduction,” is meant lowering of an event or characteristic (e.g.,tumor growth). It is understood that this is typically in relation tosome standard or expected value, in other words it is relative, but thatit is not always necessary for the standard or relative value to bereferred to. For example, “reduces tumor growth” means reducing the rateof growth of a tumor relative to a standard or a control.

By “prevent” or other forms of the word, such as “preventing” or“prevention,” is meant to stop a particular event or characteristic, tostabilize or delay the development or progression of a particular eventor characteristic, or to minimize the chances that a particular event orcharacteristic will occur. Prevent does not require comparison to acontrol as it is typically more absolute than, for example, reduce. Asused herein, something could be reduced but not prevented, but somethingthat is reduced could also be prevented. Likewise, something could beprevented but not reduced, but something that is prevented could also bereduced. It is understood that where reduce or prevent are used, unlessspecifically indicated otherwise, the use of the other word is alsoexpressly disclosed.

By “treat” or other forms of the word, such as “treated” or “treatment,”is meant to administer a composition or to perform a method in order toreduce, prevent, inhibit, or eliminate a particular characteristic orevent (e.g., tumor growth or survival). The term “control” is usedsynonymously with the term “treat.”

The term “antiviral” refers to the ability to inhibit the replication ofthe particular virus, to inhibit viral transmission, or to prevent thevirus from establishing itself in its host, and to ameliorate oralleviate the symptoms of the disease caused by the viral infection. Thetreatment is considered therapeutic if there is a reduction in viralload, decrease in mortality and/or morbidity.

The term “therapeutically effective” means the amount of the compositionused is of sufficient quantity to ameliorate one or more causes orsymptoms of a disease or disorder. Such amelioration only requires areduction or alteration, not necessarily elimination.

As used herein, the term pharmaceutically acceptable salt refers to anysalt of the compound of this invention which retains its biologicalproperties and which is not toxic or otherwise undesirable forpharmaceutical use. Such salts may be derived from a variety of organicand inorganic counterions well known in the art and include them. Suchsalts include: (1) acid addition salts formed with organic or inorganicacids such as hydrochloric, hydrobromic, sulfuric, nitric, phosphoric,sulfamic, acetic, trifluoroacetic, trichloroacetic, propionic, hexanoic,cyclopentylpropionic, glycolic, glutaric, pyruvic, lactic, malonic,succinic, sorbic, ascorbic, malic, maleic, fumaric, tartaric, citric,benzoic, 3-(4-hydroxybenzoyl)benzoic, picric, cinnamic, mandelic,phthalic, lauric, methanesulfonic, ethanesulfonic,1,2-ethane-disulfonic, 2-hydroxyethanesulfonic, benzenesulfonic,4-chlorobenzenesulfonic, 2-naphthalenesulfonic, 4-toluenesulfonic,camphoric, camphorsulfonic,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic, glucoheptonic,3-phenylpropionic, trimethylacetic, tert-butylacetic, lauryl sulfuric,gluconic, benzoic, glutamic, hydroxynaphthoic, salicylic, stearic,cyclohexylsulfamic, quinic, muconic acid and the like acids; or (2)salts formed when an acidic proton present in the parent compound either(a) is replaced by a metal ion (e.g., an alkali metal ion, an alkalineearth ion or an aluminum ion), or alkali metal or alkaline earth metalhydroxides (e.g., sodium, potassium, calcium, magnesium, aluminum,lithium, zinc, and barium hydroxide), ammonia or (b) coordinates with anorganic base, such as aliphatic, alicyclic, or aromatic organic amines,such as ammonia, methylamine, dimethylamine, diethylamine, picoline,ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine,arginine, ornithine, choline, N,N-dibenzylethylene-diamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,N-methylglucamine piperazine, tris(hydroxymethyl)-aminomethane,tetramethylammonium hydroxide, and the like. In addition, examples ofsalts include sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium and the like, and when the compound contains a basicfunctionality, salts of non-toxic organic or inorganic acids, such ashydrohalides (e.g., hydrochloride and hydrobromide), sulfate, phosphate,sulfamate, nitrate, acetate, trifluoroacetate, trichloroacetate,propionate, hexanoate, cyclopentylpropionate, glycolate, glutarate,pyruvate, lactate, malonate, succinate, sorbate, ascorbate, malate,maleate; fumarate, tartarate, citrate, benzoate,3-(4-hydroxybenzoyl)benzoate, picrate, cinnamate, mandelate, phthalate,laurate, methanesulfonate (mesylate), ethanesulfonate,1,2-ethane-disulfonate, 2-hydroxyethanesulfonate, benzenesulfonate(besylate), 4-chlorobenzenesulfonate, 2-naphthalenesulfonate,4-toluenesulfonate, camphorate, camphorsulfonate,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate, glucoheptonate,3-phenylpropionate, trimethylacetate, tert-butylacetate, lauryl sulfate,gluconate, benzoate, glutamate, hydroxynaphthoate, salicylate, stearate,cyclohexylsulfamate, quinate, muconate and the like.

The term “glycoside” or “glycosidic” compound as used herein isinterchangeable and includes reference to any of the class of compoundsthat yield a sugar and an aglycone upon hydrolysis.

The term “ANL” or “aryl naphthalene lignan” or “arylnaphthalene lignan”compound as used herein is interchangeable.

The term “aryl naphthalene lignan” or “arylnaphthalene lignan” or “ANL”as used herein includes reference to a compound comprising the basicstructure of 2,3-dimethyl-1-phenyl-naphthalene shown as below:

The carbon numbering of aryl naphthalene lignan molecule as used hereinincludes reference to a compound comprising numbering system shown asbelow:

In one class of the core structure of an aryl naphthalene compound, thetwo methyl groups are forming a γ-lactone ring to become as arylnaphthofuran-2-one lignan or aryl naphthofuran-3-one lignan shown asbelow:

The carbon numbering of an aryl naphthalene lignan glycoside molecule asused herein includes reference to a compound comprising numbering systemshown as below:

The term “hydrocarbyl” as used herein includes reference to a moietyconsisting exclusively of hydrogen and carbon atoms; such a moiety maycomprise an aliphatic and/or an aromatic moiety. The moiety may comprise1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20carbon atoms. Examples of hydrocarbyl groups include C₁₋₆ alkyl (e.g.C₁, C₂, C₃ or C₄ alkyl, for example methyl, ethyl, propyl, isopropyl.n-butyl, sec-butyl or tert-butyl); C₁₋₆ alkyl substituted by aryl (e.g.benzyl) or by cycloalkyl (e.g. cyclopropylmethyl); cycloalkyl (e.g.cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl); aryl (e.g. phenyl,naphthyl or fluorenyl); C₁₋₆ alkenyl (e.g. ethenyl, 2-propenyl or3-butenyl); C₁₋₆ alkynyl (e.g. ethynyl, 2-propynyl or 3-butynyl) and thelike.

The terms “alkyl” and “C₁₋₆ alkyl” as used herein include reference to astraight or branched chain alkyl moiety having 1, 2, 3, 4, 5 or 6 carbonatoms. This term includes reference to groups such as methyl, ethyl,propyl (n-propyl or isopropyl), butyl (n-butyl, sec-butyl ortert-butyl), pentyl, hexyl and the like. In particular, the alkyl moietymay have 1, 2, 3 or 4 carbon atoms.

The terms “alkenyl” as used herein include reference to a straight orbranched chain alkyl moiety having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms and having, in addition,at least one double bond, of either E or Z stereochemistry whereapplicable. This term includes reference to groups such as ethenyl,2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl,3-pentenyl, 1-hexenyl, 2-hexenyl and 3-hexenyl and the like.

The terms “alkynyl” as used herein include reference to a straight orbranched chain alkyl moiety having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms and having, in addition,at least one triple bond. This term includes reference to groups such asethynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl,2-pentynyl, 3-pentynyl, 1-hexynyl, 2-hexynyl and 3-hexynyl and the like.

The terms “alkoxy” and “C₁₋₆ alkoxy” as used herein include reference to—O-alkyl, wherein alkyl is straight or branched chain and comprises 1,2, 3, 4, 5 or 6 carbon atoms. In one class of embodiments, alkoxy has 1,2, 3 or 4 carbon atoms. This term includes reference to groups such asmethoxy, ethoxy, propoxy, isopropoxy, butoxy, tertbutoxy, pentoxy,hexoxy and the like.

The term “cycloalkyl” as used herein includes reference to an alicyclicmoiety having 3, 4, 5, 6, 7 or 8 carbon atoms. The group may be abridged or polycyclic ring system. More often cycloalkyl groups aremonocyclic. This term includes reference to groups such as cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, norbomyl, bicyclo[2.2.2]octyl andthe like.

The term “aryl” as used herein includes reference to an aromatic ringsystem comprising 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring carbonatoms. Aryl is often phenyl but may be a polycyclic ring system, havingtwo or more rings, at least one of which is aromatic. This term includesreference to groups such as phenyl, naphthyl, fluorenyl, azulenyl,indenyl, anthryl and the like.

“Cyclic group” means a ring or ring system, which may be unsaturated orpartially unsaturated but is usually saturated, typically containing 5to 13 ring-forming atoms, for example a 5- or 6-membered ring. Itincludes carbocyclyl and heterocyclyl moeities.

The term “carbocyclyl” as used herein includes reference to a saturated(e.g. cycloalkyl) or unsaturated (e.g. aryl) ring moiety having 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 carbon ring atoms. Inparticular, carbocyclyl includes a 3- to 10-membered ring or ring systemand, in particular, 5- or 6-membered rings, which may be saturated orunsaturated. A carbocyclic moiety is, for example, selected fromcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbomyl,bicyclo[2.2.2]octyl, phenyl, naphthyl, fluorenyl, azulenyl, indenyl,anthryl and the like.

The term “heterocyclyl” as used herein includes reference to a saturated(e.g. heterocycloalkyl) or unsaturated (e.g. heteroaryl) heterocyclicring moiety having from 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or16 ring atoms, at least one of which is selected from boron, nitrogen,oxygen, phosphorus, silicon and sulfur. In particular, heterocyclylincludes a 3- to 10-membered ring or ring system and more particularly a5 or 6-membered ring, which may be saturated or unsaturated.

A heterocyclic moiety is, for example, selected from oxiranyl, azirinyl,1.2-oxathiolanyl, imidazolyl, thienyl, furyl, tetrahydrofuryl, pyranyl,thiopyranyl, thianthrenyl, isobenzofuranyl, benzofuranyl, chromenyl,2H-pyrrolyl pyrrolyl pyrrolinyl, pyrrolidinyl, pyrrolizidinyl,imidazolyl, imidazolidinyl, benzimidazolyl, pyrazolyl pyrazinyl,pyrazolidinyl, thiazolyl, isothiazolyl, dithiazolyl, oxazolyl,isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, piperidyl, piperazinyl,pyridazinyl, morpholinyl, thiomorpholinyl, especially thiomorpholino,indolizinyl, isoindolyl, 3H-indolyl, indolyl, benzimidazolyl, cumaryl,indazolyl, triazolyl, tetrazolyl, purinyl, 4N-quinolizinyl, isoquinolyl,quinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl,octahydroisoquinolyl, benzofuranyl, dibenzofuranyl, benzothiophenyl,dibenzothiophenyl, phthalazinyl, naphthyridinyl, quinoxalyl,quinazolinyl, quinazolinyl, cinnolinyl, pteridinyl, carbazolyl,B-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl,furazanyl, phenazinyl, phenothiazinyl, phenoxazinyl, chromenyl,isochromanyl, chromanyl, 1,3,2-dioxaborolane, and the like.

The term “heterocycloalkyl” as used herein includes reference to asaturated heterocyclic moiety having 3, 4, 5, 6 or 7 ring carbon atomsand 1, 2, 3, 4 or 5 ring heteroatoms selected from nitrogen, oxygen,phosphorus and sulfur. The group may be a polycyclic ring system butmore often is monocyclic. This term includes reference to groups such asazetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, oxiranyl,pyrazolidinyl, imidazolyl, indolizidinyl, piperazinyl, thiazolidinyl,morpholinyl, thiomorpholinyl, quinolizidinyl and the like.

The term “heteroaryl” as used herein includes reference to an aromaticheterocyclic ring system having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or16 ring atoms, at least one of which is selected from nitrogen, oxygenand sulfur. The group may be a polycyclic ring system, having two ormore rings, at least one of which is aromatic, but is more oftenmonocyclic. This term includes reference to groups such as pyrimidinyl,furanyl, benzobthiophenyl, thiophenyl, pyrrolyl, imidazolyl,pyrrolidinyl, pyridinyl, benzobfuranyl, pyrazinyl, purinyl, indolyl,benzimidazolyl, quinolinyl, phenothiazinyl, triazinyl, phthalazinyl,2H-chromenyl, oxazolyl, isoxazolyl, thiazolyl, isoindolylindazolyl,purinyl, isoquinolinyl, quinazolinyl, pteridinyl and the like.

The term “halogen” as used herein includes reference to F, Cl, Br or I.

The expression “halogen containing moiety” as used herein includesreference to a moiety comprising 1 to 30 plural valence atoms selectedfrom carbon, nitrogen, oxygen and sulfur which moiety includes at leastone halogen. The moiety may be hydrocarbyl for example C₁₋₆ alkyl orC₁₋₆ alkoxy, or carbocyclyl for example aryl.

The term “substituted” as used herein in reference to a moiety meansthat one or more, especially up to 5, more especially 1, 2 or 3, of thehydrogen atoms in said moiety are replaced independently of each otherby the corresponding number of the described substituents. The term“optionally substituted as used herein means substituted orun-substituted. It will, of course, be understood that substituents areonly at positions where they are chemically possible, the person skilledin the art being able to decide (either experimentally or theoretically)without inappropriate effort whether a particular substitution ispossible.

Where two or more moieties are described as being “each independently”selected from a list of atoms or groups, this means that the moietiesmay be the same or different. The identity of each moiety is thereforeindependent of the identities of the one or more other moieties.

The term “enantiomer” as used herein means one of two stereoisomers thathave mirror images of one another.

The term “stereoisomer” as used herein means one of class of isomericmolecules that have the same molecular formula and sequence of bondedatoms, but different three-dimensional orientations of their atoms inspace.

The term “tautomer” means isomeric molecules that readily interconvertby a chemical reaction. The reaction commonly results in the migrationof a hydrogen atom, which results in a switch of a single bond andadjacent double bond.

A prodrug is a medication that is administered as an inactive (or lessthan fully active) chemical derivative that is subsequently converted toan active pharmacological agent in the body, often through normalmetabolic processes.

CC₅₀ is a cytotoxicity measure of the concentration for a test drug toinhibit cell growth by 50%.

EC₅₀ is an antiviral activity measure of the effective concentration fora test drug to inhibit viral growth by 50%.

The term “selectivity index” or “SI” means a ratio that measures thewindow between cytotoxicity and antiviral activity by dividing the givenCC₅₀ value into the IC₅₀ value (CC₅₀/IC₅₀) of a test drug. The higher SIratio means more effective and safer a test drug would be for a givenviral infection in an in vitro experiment.

The symbol

in a chemical structure represents a position from where the specifiedchemical structure is bonded to another chemical structure.

The symbol “β” in a chemical structure indicates that the bondconnection is above (or before) the plane of the paper or screen. Thesymbol “α” in a chemical structure indicates that the bond connection isbelow (or behind) the plane of the paper or screen.

A solid wedge in a chemical structure indicates that this bond is above(or before) the plane of the paper or screen toward to the viewer. Ahashed (or broken) wedge in a chemical structure indicates that the bondconnection is below (or behind) the plane of the paper or screenreceding away from the viewer.

Provided herein is a compound and use of the the compound in themanufacture of a medicament for treating a viral infection and a methodof using the compound for treating a viral infection in a subject,comprising the step of administering a therapeutically effective amountof the compound to the subject, wherein the compound has the Formula(I):

or a pharmaceutically acceptable salt or pro-drug thereof wherein,

X is oxygen or sulfur;

R¹ is R¹⁵, —OR¹⁵, —C(O)R¹⁵, or —C(O)OR¹⁵;

R², R⁵, R⁶, R¹⁰, R¹³, and R¹⁴ are each independently hydrogen orhalogen;

R³ and R⁴ are each independently selected from the group consisting of—OR^(D) and —OC(O)R¹⁵; or R³ and R⁴ taken together with the carbon atomsto which they are attached form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR⁶;

R⁷, R⁸, and R⁹ are each independently selected from the group consistingof —OR¹⁵ and —OC(O)R¹⁵; or R⁷ and R⁸ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶; or R⁸ and R⁹ taken together with the carbon atoms towhich they are attached to form a 5-6 membered heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR⁶;

R¹¹ and R¹² taken together form oxo; or while one of R¹¹ and R¹² ishydrogen or halogen, the other one of R¹¹ and R¹² is selected from thegroup consisting of R'S, —OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹⁵ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, heteroaryl, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N═C(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸,—N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸, —N(R¹⁷)S(O)₂R¹⁸, 1,3,2-dioxaborolaneoptionally substituted with 1, 2, 3, or 4 group(s) independentlyselected from alkyl, a glycosidic group, alkynyl optionally substitutedwith a trialkylsilane, hydrocarbyl optionally substituted with 1, 2, 3,4 or 5 group(s) independently selected from R¹⁶, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from R¹⁶, and —(CH₂)_(k)-heterocyclyl optionally substitutedwith 1, 2, 3, 4 or 5 group(s) independently selected from R¹⁶, wherein kis an integer between 1-6;

R¹⁶ for each occurrence is independently selected from the groupconsisting of alkynyl, halogen, trichloromethyl, trifluoromethyl, cyano,nitro, oxo, ═NR¹⁷, —OR¹⁷, —C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷,—OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸,—N(R¹⁷)C(O)R¹⁸ and —N(R¹⁷)S(O)₂R¹⁸; and

R¹⁷ and R¹⁸ for each occurrence are independently hydrogen, alkyl,alkynyl, cycloalkyl, aryl, or heteroaryl, or selected from hydrocarbyland heterocyclyl, either of which is optionally substituted with 1, 2,3, 4 or 5 group(s) independently selected from the group consisting ofhalogen, trichloromethyl, trifluoromethyl, cyano, amino, hydroxy, C₁₋₆alkyl and C₁₋₆ alkoxy.

In certain embodiments, the compound of Formula (I) is not a compoundselected from the group consisting of 5, 15a, 15b, 16, 17b, 19d,25a-25g, and 26a-26g:

In certain embodiments, R¹ is hydrogen, alkyl, aryl, heteroaryl (such asfuran, thiophene, pyridine, and the like), —N(R¹⁷)R¹⁸, —N═C(R¹⁷)(R¹⁸),alkynyl optionally substituted with a trialkylsilane, a glycosidicgroup, 1,3,2-dioxaborolane optionally substituted with 1, 2, 3, or 4group(s) independently selected from alkyl, or —O—(CR₂)_(m)—R¹⁶, whereinm is a whole number selected from 1-4, 1-3, or 1-2; R for eachoccurrence is independently hydrogen, alkyl, cycloalkyl, or aryl; andR¹⁶ is alkynyl, cyano, —OR¹⁷, —N(R¹⁷)R¹⁸, —C(O)N(R¹⁷)R¹⁸, or —C(O)OR¹⁷.

In certain embodiments, R¹ is —O—CH₂—R¹⁶, wherein R¹⁶ is —C≡CH, cyano,—OR¹⁷, —N(R¹⁷)R¹⁸, —C(O)N(R¹⁷)R¹⁸, or —C(O)OR¹⁷.

In certain embodiments, each of R³ and R⁴ is independently selected fromthe group consisting of —OR¹⁵ and —OC(O)R¹⁵, wherein R¹⁵ is hydrogen,alkyl, cycloalkyl, heterocyclcyl, aryl, and heteroaryl; or R³ and R⁴taken together with the carbon atoms to which they are attached form a5-6 membered heterocyclyl. In certain embodiments, R³ and R⁴ is —OR¹⁵,wherein R¹⁵ is C₁-C₆ alkyl, C₁-C₄ alkyl, or C₁-C₂ alkyl.

In certain embodiments, each of R⁸ and R⁹ is independently selected fromthe group consisting of —OR¹⁵ and —OC(O)R¹⁵, wherein R¹⁵ is hydrogen,alkyl, cycloalkyl, heterocyclcyl, aryl, and heteroaryl; or R⁸ and R⁹taken together with the carbon atoms to which they are attached form a5-6 membered heterocyclyl. In certain embodiments, R⁸ and R⁹ takentogether with the carbon atoms to which they are attached form amethylenedioxy ring.

In certain embodiments, R¹¹ and R¹² taken together form oxo.

In certain embodiments, each of R³ and R⁴ is —O-alkyl; R⁸ and R⁹ takentogether with the carbon atoms to which they are attached form amethylenedioxy ring; R¹ is heteroaryl, —OR¹⁵, —C(O)R¹⁵, —N(R¹⁷)R¹⁸,—N(R¹⁷)C(O)R¹⁸, —N═C(R¹⁷)R¹⁸, pinacolboryl, —OS(O)₂CF₃, a glycosidicgroup, heterocyclcyl optionally substituted with 1 or 2 group(s)independently selected from R¹⁶ or, alkynyl optionally substituted witha trialkylsilane; or R¹ is —OCH₂-cyano, —OCH₂—C≡CH, —OCH₂—C(O)N(R¹⁷)R¹⁸,or —C(O)OR¹⁷; and R¹¹ and R¹² taken together form oxo.

In certain embodiments, the compound is selected compounds 12a, 12b, 13,14a, 14b, 17a, 17c, 17d, 17e, 17f, 17g, 17h, 18, 19a, 19b, 19c, 19e,19f, 19g, 19h, 19i, 19j, 19k, 19l, 19m, 19n, 19o, 19p, 20, 21, 22, 23and 24:

In certain embodiments, R¹ is a glycosidic group represented by theFormula (V):

wherein, R¹⁹ and R²⁰ taken together to form oxo; or while one of R¹⁹ andR²⁰ is hydrogen or halogen, the other one of R¹⁹ and R²⁰ is selectedfrom the group consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷;

R²¹ and R²² taken together to form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R²¹ and R²² is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²⁰ and R²² taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl, which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy;

R²³ and R²⁴ taken together to form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; or R²² and R²⁴ taken together with the carbon atoms to whichthey are attached to form a 5-6 membered heterocyclyl, which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy;

R²⁵ and R²⁶ taken together form oxo; or while one of R²⁵ and R²⁶ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from thegroup consisting of R²⁷, —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)OR²⁷,—CH₂R²⁷, and —C(O)R²⁷; or R²⁴ and R²⁶ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyl,which is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyland C₁₋₆ alkoxy;

R²⁷ for each occurrence is independently selected from the groupconsisting of hydrogen, halogen, trichloromethyl, trifluoromethyl,cyano, nitro, —OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹,—S(O)₂R²⁹, —S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰,—N(R²⁹)C(O)R³⁰, —N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with1, 2, 3, 4 or 5 group(s) independently selected from the groupconsisting of R²⁸, heterocyclcyl optionally substituted with 1, 2, 3, 4or 5 group(s) independently selected from the group consisting of R²⁸,and —(CH₂)_(k)-heterocyclyl optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from the group consisting of R²⁸,wherein k is an integer between 1-6;

R²⁸ for each occurrence is independently selected from halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰ and—N(R²⁹)S(O)₂R³⁰; and

R²⁹ and R³⁰ for each occurrence are each independently hydrogen orselected from hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy.

In certain embodiments, the glycosidic group is a monosaccharide,disaccharide, trisaccharide, tetrasaccharide or polysaccharide group,and may exist in any isomeric form, for example α-D, α-L, β-D or β-Lforms. In certain embodiments, the glycloside is a monosaccharideselected from the group consisting of an α-L isomer and an β-L isomer.

In certain embodiments, R¹⁹, R²¹, R²³, and R²⁵ are each hydrogen; R²⁰,R²², and R²⁴ are each independently selected from the group consistingof —OR²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)R²⁷ and —OC(O)OR²⁷; and R²⁶ ishydrogen, methyl, —OR²⁷, —OC(O)R²⁷, or —CH₂—OC(O)R²⁷; or R²⁰ and R²²taken together with the carbon atoms to which they are attached form a5-6 membered heterocyclyl, which is optionally substituted with 1, 2, 3,4 or 5 group(s) independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy;or R²² and R²⁴ taken together with the carbon atoms to which they areattached form a 5-6 membered heterocyclyl, which is optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromC₁₋₆ alkyl and C₁₋₆ alkoxy; or R²⁴ and R²⁶ taken together with thecarbon atoms to which they are attached form a 5-6 memberedheterocyclyl, which is optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from C₁₋₆ alkyl and C₁₋₆ alkoxy; and theglycloside is a monosaccaride.

In certain embodiments, R²⁰, R²², and R²⁴ are each independentlyselected from the group consisting of —OR²⁷, —OC(O)N(R²⁷)R²⁷, —OC(O)R²⁷and —OC(O)OR²⁷, wherein R²⁷ for each occurrence is independentlyselected from the group consisting of hydrogen, alkyl, cycloalkyl,alkenyl, aryl, aralkyl, heteroaryl, perhaloalkyl, and —C(O)CH₂CN.

In instances in which R¹ is a glycosidic group, the compound can beselected from the group consisting of: 27aa, 27ab, 27ac, 27ad, 27ae,27af, 27ba, 27bb, 27bc, 27bd, 27be, 27bf, 28ab1, 28ab2, 28ab3, 28bb1,28bb2, 28bb3, 29a, 29b, 30a, 30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i,30j, 30k, 30l, 31a, 31b, 31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, 31k,31l, 32a, 32b, 32c and 32d:

The present disclosure also provides aryl naphthalene glycosidecompounds, including derivatives of diphyllin, patentiflorin A,justiprocuminA or justiprocumin B useful in the treatment of viralinfections.

In certain embodiments, the compound has the Formula (I) or (II):

or a stereoisomer thereof; or an enantiomer thereof; or apharmaceutically acceptable salt or pro-drug thereof wherein

X is oxygen or sulfur;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹ and R¹⁰ are each independentlyhydrogen, halogen or a moiety comprising 1 to 30 plural valence atomsselected from the group consisting of carbon, nitrogen, oxygen andsulfur; or R² and R³, R³ and R⁴, R⁴ and R⁵, R⁶ and R⁷, R⁷ and R⁸, R⁸ andR⁹ or R⁹ and R¹⁰ may be taken together with the carbon atoms to whichthey are attached to form a cyclic group which is optionally substitutedwith halogen and a moiety comprising 1 to 30 plural valence atomsselected from the group consisting of carbon, nitrogen, oxygen andsulfur;

R¹¹ and R¹² taken together form oxo; or while one of R¹¹ and R¹² ishydrogen or halogen, the other one of R¹¹ and R¹² is selected from thegroup consisting of R¹⁵, —OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹³ and R¹⁴ taken together form oxo; or while one of R¹³ and R¹⁴ ishydrogen or halogen, the other one of R¹³ and R¹⁴ is selected from thegroup consisting of R¹⁵, —OR¹⁵, —C(O)R¹⁵ and —C(O)OR¹⁵;

R¹⁵ is independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, hydrocarbyloptionally substituted with 1, 2, 3, 4 or 5 R¹⁶, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 R¹⁶, —(CH₂)_(k)-heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 R¹⁶, —OR¹⁷, —C(O)R¹⁸,—C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷, —OC(O)R¹⁷, —S(O)₂R¹⁷, —S(O)₂N(R¹⁷)R¹⁸,—N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸ and —N(R¹⁷)S(O)₂R¹⁸,wherein k is an integer between 1 and 6 (e.g. 1, 2 or 3);

R¹⁶ is independently selected from the group consisting of halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR¹⁷, —OR¹⁷,—C(O)R¹⁸, —C(O)N(R¹⁷)R¹⁸, —C(O)OR¹⁷, —OC(O)R¹⁷, —S(O)₂R¹⁷,—S(O)₂N(R¹⁷)R¹⁸, —N(R¹⁷)R¹⁸, —N(R¹⁷)N(R¹⁷)R¹⁸, —N(R¹⁷)C(O)R¹⁸ and—N(R¹⁷)S(O)₂R¹⁸; and

R¹⁷ and R¹⁸ are each independently hydrogen or selected from the groupconsisting of hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 substituents independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy.

In certain embodiments, the compounds has the Formula (III) or (IV):

or a stereoisomer thereof; or an enantiomer thereof; of apharmaceutically acceptable salt or pro-drug thereof wherein,

X is sulfur or oxygen;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸ and R⁹ are each independently hydrogen,halogen or a moiety comprising 1 to 30 plural valence atoms selectedfrom carbon, nitrogen, oxygen and sulfur; or R¹ and R², R² and R³, R³and R⁴, R⁵ and R⁶, R⁶ and R⁷, R⁷ and R⁸, and/or R⁸ and R⁹ taken togetherwith the carbon atoms to which they are attached to form a cyclic groupwhich is optionally substituted with 1, 2, 3, 4 or 5 group(s) selectedfrom halogen and a moiety comprising 1 to 30 plural valence atomsselected from the group consisting of carbon, nitrogen, oxygen andsulfur;

R¹⁰ and R¹¹ taken together form oxo; or while one of R¹⁰ and R¹¹ ishydrogen or halogen, the other one of R¹⁰ and R¹¹ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, and —C(O)OR²⁷;

R¹² and R¹³ taken together form oxo; or while one of R¹² and R¹³ ishydrogen or halogen, the other one of R¹² and R¹³ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, and —C(O)OR²⁷;

R¹⁹ and R²⁰ taken together form oxo; or while one of R¹⁹ and R²⁰ ishydrogen or halogen, the other one of R¹⁹ and R²⁰ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²¹ and R²² taken together form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R¹⁶ and R¹⁷ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide andsubstituted tetrasaccharide;

R²³ and R²⁴ taken together form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²⁵ and R²⁶ taken together form oxo; or while one of R²⁵ and R²⁶ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, —CH₂R³¹ and—C(O)R³¹;

R²⁷ is independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰,—N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with 1, 2, 3, 4 or 5group(s) selected from the group consisting of R²³, heterocyclcyloptionally substituted with 1, 2, 3, 4 or 5 group(s) selected from thegroup consisting of R²⁸, and —(CH₂)_(k)-heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) selected from group(s)independently selected R²⁸, wherein k is an integer between 1 and 6(e.g. 1, 2 or 3);

R²⁸ is independently selected from the group consisting of halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰ and—N(R²⁹)S(O)₂R³⁰;

R²⁹ and R³⁰ are each independently hydrogen or selected from the groupconsisting of hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy;

R³¹ is independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰,—N(R²⁹)S(O)₂R³⁰, monosaccharide, substituted monosaccharide,disaccharide, substituted disaccharide, trisaccharide, substitutedtrisaccharide, tetrasaccharide, substituted tetrasaccharide, hydrocarbyloptionally substituted with 1, 2, 3, 4 or 5 group(s) selected from thegroup consisting of R²⁸, and heterocyclyl optionally substituted with 1,2, 3, 4 or 5 group(s) selected from the group consisting of R²⁸; and

R is hydrogen or selected from hydrocarbyl and heterocyclyl, either ofwhich is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyland C₁₋₆ alkoxy.

In certain embodiments, the glycosidic group can be a carbohydrategroup, such as a monosaccharide, disaccharide, trisaccharide,tetrasaccharide or polysaccharide group, and may exist in any isomericform, for example α-D, α-L, β-D or β-L forms. The carbohydrate group maybe optionally substituted with other type of substituents or evenadditional glycosidic groups. However, the total number ofmonosaccharide and substituted monosaccharide contained in the chemicalstructure of a compound is generally 30 or less.

In certain embodiments, the glycosidic group may comprise one or moremonosaccaride units of Formula (V) or (VI):

wherein R¹⁹ and R²⁰ taken together form oxo; or while one of R¹⁹ and R²⁰is hydrogen or halogen, the other one of R¹⁹ and R²⁰ is selected fromthe group consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²¹ and R²² taken together form oxo; or while one of R²¹ and R²² ishydrogen or halogen, the other one of R²¹ and R²² is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide, andsubstituted tetrasaccharide;

R²³ and R²⁴ taken together form oxo; or while one of R²³ and R²⁴ ishydrogen or halogen, the other one of R²³ and R²⁴ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, monosaccharide,substituted monosaccharide, disaccharide, substituted disaccharide,trisaccharide, substituted trisaccharide, tetrasaccharide andsubstituted tetrasaccharide;

R²⁵ and R²⁶ taken together form oxo; or while one of R²⁵ and R²⁶ ishydrogen or halogen, the other one of R²⁵ and R²⁶ is selected from thegroup consisting of R²⁷, —OR²⁷, —C(O)R²⁷, —C(O)OR²⁷, —CH₂R³¹, and—C(O)R³¹;

R²⁷ is independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰,—N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from the group consisting of R²⁸,heterocyclcyl optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from the group consisting of R²⁸, and—(CH₂)_(k)-heterocyclyl optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from the group consisting of R²⁸,wherein k is an integer between 1 and 6 (e.g. 1, 2 or 3);

R²⁸ is independently selected from the group consisting of halogen,trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰ and—N(R²⁹)S(O)₂R³⁰;

R²⁹ and R³⁰ are each independently hydrogen or selected from the groupconsisting of hydrocarbyl and heterocyclyl, either of which isoptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyl and C₁₋₆alkoxy; and

R³¹ is independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰,—N(R²⁹)S(O)₂R³⁰, monosaccharide, substituted monosaccharide,disaccharide, substituted disaccharide, trisaccharide, substitutedtrisaccharide, tetrasaccharide, substituted tetrasaccharide, hydrocarbyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from the group consisting of R²⁸, heterocyclcyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromthe group consisting of R²⁸, and —(CH₂)_(k)-heterocyclyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromthe group consisting of R²⁸, wherein k is an integer between 1 and 6(e.g. 1, 2 or 3).

In certain embodiments, the glycosidic group comprises 1, 2, 3, or 4monosaccaride units, wherein each monosaccharide unit in the glycosidicgroup is independently for each occurrence a monosaccaride unit ofFormula (V) or Formula (VI).

In certain embodiments, R¹, R⁴, R⁵, R⁸ and R⁹ are each independentlyhydrogen, hydroxy or methoxy; R² and R³ are each independently hydrogen,hydroxy, methoxy, or taken together with the carbon atoms to which theyare attached form a [1,3] dioxole cyclic group; R⁶ and R⁷ are eachindependently hydrogen, hydroxy, methoxy; or R⁶ and R⁷ taken togetherwith the carbon atoms to which they are attached form a [1,3] dioxolecyclic group; R¹⁰ and R¹¹ taken together form oxo; R¹² and R¹³ are eachindependently hydrogen; and X is oxygen.

Exemplary glycosidic groups include glucopyranoside, galactopyranoside,mannopyranoside, fucopyranoside, arabinopyranoside, glucopyranoside,galactopyranoside, glucuronide, lactopyranoside, xylopyranoside,glucosaminide, galactosaminide, alloside, lyxoside, taloside, threoside,riboside, fructoside, rhamnoside and guloside groups. More particularly,the glycosidic group may be selected from α-D-glucopyranoside,α-D-galactopyranoside, α-D-mannopyranoside, α-L-fucopyranoside,α-L-arabinopyranoside, β-D-glucopyranoside, β-D-galactopyranoside,β-D-glucuronide, β-D-lactopyranoside, β-D-xylopyranoside,β-D-glucosaminide, β-D-galactosaminide, β-D-alloside, β-D-lyxoside,β-D-taloside, β-D-threoside, β-D-riboside, β-D-fructoside,β-D-rhamnoside and β-L-guoside groups.

Examples of compounds of the present disclosure include those shownbelow. It will of course be appreciated that, where appropriate, eachcompound may be in the form of the free compound, an acid or baseaddition salt, or a prodrug.

The present disclosure also provides a pharmaceutical compositioncomprising at least one of the compounds described herein and at leastone pharmaceutically acceptable excipient.

The compounds described herein and their pharmaceutically acceptablesalts can be administered to a subject either alone or in combinationwith pharmaceutically acceptable, excipients, carriers, and/or diluentsin a pharmaceutical composition according to standard pharmaceuticalpractice. The compounds can be administered orally or parenterally.Parenteral administration includes intravenous, intramuscular,intraperitoneal, subcutaneous and topical, the preferred method beingintravenous and topical administrations.

Accordingly, the present disclosure provides pharmaceutically acceptablecompositions, which comprise a therapeutically effective amount of oneor more of the compounds described herein, formulated together with oneor more pharmaceutically, excipients, acceptable carriers (additives)and/or diluents. The pharmaceutical compositions of the presentdisclosure may be specially formulated for administration in solid orliquid form, including those adapted for the following: (1) parenteraladministration, for example, by subcutaneous, intramuscular, intravenousor epidural injection as, for example, a sterile solution or suspension,or sustained-release formulation; and (2) oral administration, forexample, drenches (aqueous or non-aqueous solutions or suspensions),tablets, e.g., those targeted for buccal, sublingual, and systemicabsorption, boluses, powders, granules, pastes for application to thetongue.

As set out herein, certain embodiments of the compounds described hereinmay contain a basic functional group, such as amino, and are, thus,capable of forming pharmaceutically acceptable salts withpharmaceutically acceptable acids. The term “pharmaceutically acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present disclosure.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting a purifiedcompound of the invention in its free base form with a suitable organicor inorganic acid, and isolating the salt thus formed during subsequentpurification. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, nitrate, acetate, valerate, oleate,palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate,glucoheptonate, lactobionate, and laurylsulphonate salts and the like.

The pharmaceutically acceptable salts of the compounds of the presentdisclosure include the conventional non-toxic salts or quaternaryammonium salts of the compounds, e.g., from non-toxic organic orinorganic acids. For example, such conventional non-toxic salts includethose derived from inorganic acids such as hydrochloride, hydrobromic,sulfuric, sulfamic, phosphoric, nitric, and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isothionic, and the like.

In other cases, the compounds described herein may contain one or moreacidic functional groups and, thus, are capable of formingpharmaceutically acceptable salts with pharmaceutically acceptablebases. The term “pharmaceutically acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present disclosure. These salts can likewisebe prepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting the purified compoundin its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically-acceptable metal cation,with ammonia, or with a pharmaceutically-acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives, solubilizing agents, buffers and antioxidants can also bepresent in the compositions.

Methods of preparing these formulations or compounds include the step ofbringing into association a compound described herein with the carrieror excipient and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present disclosure withliquid carriers (liquid formulation), liquid carriers followed bylyophylization (powder formulation for reconstitution with sterile wateror the like), or finely divided solid carriers, or both, and then, ifnecessary, shaping or packaging the product.

Pharmaceutical compositions of the present disclosure suitable forparenteral administration comprise one or more compounds describedherein in combination with one or more pharmaceutically-acceptablesterile isotonic aqueous or non-aqueous solutions, dispersions,suspensions or emulsions, or sterile powders which may be reconstitutedinto sterile injectable solutions or dispersions just prior to use,which may contain sugars, alcohols, antioxidants, buffers,bacteriostats, chelating agents, solutes which render the formulationisotonic with the blood of the intended recipient or suspending orthickening agents.

Examples of suitable aqueous and non-aqueous carriers which may beemployed in the pharmaceutical compositions of the disclosure includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants, such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms upon the compounds of the presentdisclosure may be ensured by the inclusion of various antibacterial andantifungal agents, for example, paraben, chlorobutanol, phenol sorbicacid, and the like. It may also be desirable to include isotonic agents,such as sugars, sodium chloride, and the like into the compositions. Inaddition, prolonged absorption of the injectable pharmaceutical form maybe brought about by the inclusion of agents which delay absorption, suchas aluminum monostearate and gelatin.

The present disclosure provides diphyllin and patentiflorin A analogswith anti-HIV and anti-influenza virus activity and synthesis thereof.The compounds of the present disclosure were synthesized and evaluatedfor their anti-HIV and anti-influenza viral activity.

The synthetic or semi-synthetic analogues of diphyllin can beexemplified by 12a, 12b, 13, 14a, 14b, 15a, 15b, 16, 17a, 17b, 17c, 17d,17e, 17f, 17g, 17h, 18, 19a, 19b, 19c, 19d, 19e, 19f, 19g, 19h, 19i,19j, 19k, 19l, 19m, 19n, 19o, 19p, 20, 21, 22, 23 and 24.

The synthetic or semi-synthetic analogues of patentiflorin A can beexemplified by 25a, 25b, 25c, 25d, 25e, 25f, 25g, 26a, 26b, 26c, 26d,26e, 26f, 26g, 27aa, 27ab, 27ac, 27ad, 27ae, 27af, 27ba, 27bb, 27bc,27bd, 27be, 27bf, 28ab1, 28ab2, 28ab3, 28bb1, 28bb2, 28bb3, 29a, 29b,30a, 30b, 30c, 30d, 30e, 30f, 30g, 30h, 30i, 30j, 30k, 30l, 31a, 31b,31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, 31k, 31l, 32a, 32b, 32c and 32d.

EXAMPLES

The following examples are set forth below to illustrate the methods andresults according to the disclosed subject matter. These examples arenot intended to be inclusive of all aspects of the subject matterdisclosed herein, but rather to illustrate representative methods,compositions, and results. These examples are not intended to excludeequivalents and variations of the present invention, which are apparentto one skilled in the art.

Unless indicated otherwise, parts are parts by weight, temperature is °C. or is at ambient temperature, and pressure is at or near atmospheric.There are numerous variations and combinations of reaction conditions,e.g., component concentrations, temperatures, pressures, and otherreaction ranges and conditions that can be used to optimize the productpurity and yield obtained from the described process. Only reasonableand routine experimentation will be required to optimize such processconditions.

Examples of compounds of the present disclosure include those shownbelow. It will of course be appreciated that, where appropriate, eachcompound may be in the form of the free compound, an acid or baseaddition salt, or a prodrug.

By using diphyllin (5) as a structural scaffold, the general syntheticroute for its C—O derivatization on C-7 is illustrated in FIG. 2 .Esters 12a and 12b were prepared from 5 by a substitution reaction.Acidic hydrolysis of ester 12a provided acid 13, which was converted toamides 14a and 14b by coupling with ammonium chloride (NH₄Cl) or anamine in the presence of1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU) or 1-hydroxybenzotriazole hydrate(HOBt). O-alkoxyl alcohols 15a and 15b were obtained from the reactionsof 5 with the corresponding epoxides ethylene oxide and isobutyleneoxide, respectively. Derivatives 17a-17h were prepared from 5,respectively, in one step. Alkylation of 5 with cesium carbonate(Cs₂CO₃) followed by bromoacetonitrile, iodomethane or propargyl bromiderendered derivatives 17a-17c, respectively. Addition of potassiumcarbonate (K₂CO₃) and 2-chloropyrimidinein to 5 in dimethylformamide(DMF) under 100° C. provided compound 17d. Similarly, the addition oftrimethylamine (Me₃N), 4-dimethylaminopyridine (DMAP) and correspondingacyl chlorides to 5 gave esters 17e-17h, respectively. The synthesis forC-7 alkyl derivatives (FIG. 3 ) envisioned conversion of 5 to thecorresponding trifluoromethanesulfonate 18, which would undergopalladium-catalyzed coupling reactions to afford derivatives 19a-19p and20-24.

ANL analogs and compounds 25a-25g and 26a-26g containing different sugarunits outlined in FIG. 4 were prepared, and which were further modifiedto provide additional ANL analogs 27aa-27bf, 28ab1-28bb3, 29a, 29b,30a-30l and 31a-30l outlined in FIG. 5 .

Glycosylation of diphyllin (5) at C-7 with different sugar units (a-g)led to synthesis of a series of diphyllin glycosides (26a-26g),respectively (FIG. 4 ). The synthesis involved the production ofglycosyl bromides through bromination reactions of the per-acetylatedglycosides, followed by its phase transfer catalysis (PTC) glycosylationtreatment with 0.1 mol/L aqueous sodium hydroxide (NaOH) in the presenceof tetrabutylammonium bromide (TBAB) at 40° C. The resulted per-acetylglycosides 25a-25g were subsequently deacetylated with K₂CO₃ in methanol(MeOH) to afford the deprotected glycosides 26a-26g, respectively.

To significantly increase the number of the ANL library compounds,further structural modification on the hydroxy groups in the sugar unitsof ANLs (FIG. 5 ) were proposed. Firstly, 3″-O-acetyl diphyllinglycosides 27aa-27af and 27ba-27bf were prepared from the reactions ofacetic anhydride (Ac₂O) and tetrabutylammonium acetate (TBOAc) with thecorresponding 26a and 26b. Separation by silica gel columnchromatography of the reaction mixture of 26a afforded the six singlecompounds 27aa-27af (27aa, 27ab, 27ac, 27ad, 27ae and 27af). Separationby silica gel column chromatography of the reaction mixture of 26bafforded the six single compounds 27ba-27bf (27ba, 27bb, 27bc, 27bd,27be and 27bf).

Compound 27ab or 27bb was treated with allyl chloroformate (AllocOCl) togive the bis-protected ANL glycosides the corresponding 28ab1-28ab3 and28bb1-28bb3. Separation by silica gel column chromatography of thereaction mixture of 27ab afforded the three single compounds 28ab1-28ab3(28ab1, 28ab2 and 28ab3). Separation by silica gel column chromatographyof the reaction mixture of 27bb afforded the three single compounds28bb1-28bb3 (28bb1, 28bb2 and 28bb3). The acetyl group in compound 28ab3or 28bb3 was removed by further treatment with acetyl chloride (AcCl) togive the bis-choloroformate protected ANL glycosides 29a and 29b,respectively. Substitution of 29a or 29b with triethylamine (Et₃N) andDMAP, followed by different acyl chlorides yielded derivatives 30a-k,respectively. Addition of potassium hydroxide (KOH), 18-crown-6 andbenzyl bromide to 29a in tetrahydrofuran (THF) under r.t. providedcompound 30l. The two allyl groups of 30a-l were deprotected bytreatment of tetrakis(triphenylphosphine)palladium(0)[Pd(PPh₃)₄],triphenylphosphine (PPh₃), Et₃N and HCOOH in THF to produce derivatives31a-l, respectively.

The compounds described herein can exhibit broad antiviral properties.According to methods of the present disclosure, compounds of Formula (I)are administered to a patient to inhibit replication of or reducecytopathic effects of viruses, such as IV, coronaviruses, Ebola virus,Marburg virus or influenza viruses. Other viruses that may be inhibitedby compounds of Formula (I) include, but are not limited to,cytomegalovirus (CMV), HSV-1 (herpes simplex virus type 1), HSV-2(herpes simplex virus type 2), HBV (hepatitis B virus), HCV (hepatitis Cvirus), HPV (human papilloma virus), influenza A, influenza B, RSV(respiratory syncitial virus), RV (rhinovirus), AV (adenovirus), PIV(human parainfluenza viruses), Epstein-Barr virus (EBV), varicellazoster virus (VZV), dengue virus and Zika virus.

The compounds, pharmaceutical compositions, and therapeutic methodsdescribed herein are useful for preventing or treating or amelioratingHIV infections.

The compounds, pharmaceutical compositions, and therapeutic methodsdescribed herein are useful for preventing, treating, or amelioratinginfections caused by influenza viruses, including but not limited to:any of the subtypes of influenza A, influenza B, or influenza C.

In certain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing,treating, or ameliorating infections caused by influenza A viruses,including but not limited to, any of the strains of H1N1, H1N2, H1N3,H1N4, H1N5, H1N6, H1N7, H1N8, H1N9, H2N1, H2N2, H2N3, H2N4, H2N5, H2N6,H2N7, H2N8, H2N9, H3N1, H3N2, H3N3, H3N4, H3N5, H3N6, H3N7, H3N8, H3N9,H4N1, H5N1, H5N2, H5N3, H5N4, H5N5, H5N6, H5N7, H5N8, H5N9, H6N1, H6N2,H6N3, H6N4, H6N5, H6N6, H6N7, H6N8, H6N9, H7N1, H7N2, H7N3, H7N4, H7N5,H7N6, H7N7, H7N8, H7N9, H8N1, H8N2, H8N3, H8N4, H8N5, H8N6, H8N7, H8N8,H8N9, H9N1, H9N2, H9N3, H9N4, H9N5, H9N6, H9N7, H9N8, H9N9, H10N1,H10N2, H10N3, H10N4, H10N5, H10N6, H10N7, H10N8, H10N9, H11N1, H11N2,H11N3, H11N4, H11N5, H11N6, H11N7, H11N8, H11N9, H12N1, H12N2, H12N3,H12N4, H12N5, H12N6, H12N7, H12N8, H12N9, H13N1, H13N2, H13N3, H13N4,H13N5, H13N6, H13N7, H13N8, H13N9, H14N1, H14N2, H14N3, H14N4, H14N5,H14N6, H14N7, H14N8, H14N9, H15N1, H15N2, H15N3, H15N4, H15N5, H15N6,H1N7, H15N8, and H15N9.

In certain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing,treating, or ameliorating infections caused by influenza A virus strainshaving a type 5 hemagglutinin protein. In certain embodiments, theinfluenza A virus strain has a type 5 hemagglutinin protein andneuraminidase protein selected from types 1 to 11. In certainembodiments, the influenza A virus is selected from the group consistingof H5N1 and H5N2.

In certain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing,treating, or ameliorating infections caused by H5N1.

The term “HIV”, as used herein, refers to the human immunodeficiencyvirus and includes HIV-1, HIV-2 and SIV. In certain embodiments, HIVrefers to HIV-1 and/or HIV-2. “HIV-1” means the human immunodeficiencyvirus type-1. HIV-1 can include but is not limited to extracellularvirus particles and the forms of HIV-1 associated with HIV-1 infectedcells. The HIV-1 virus can include any of the known major subtypes(classes A, B, C, D, E, F, G and H) or outlying subtype (group O)including laboratory strains and primary isolates. “HIV-2” means thehuman immunodeficiency virus type-2. HIV-2 can include but is notlimited to extracellular virus particles and the forms of HIV-2associated with HIV-2 infected cells. The term “SIV” refers to simianimmunodeficiency virus, which is an HIV-like virus that infects monkeys,chimpanzees, and other nonhuman primates. SIV can include but is notlimited to extracellular virus particles and the forms of SIV associatedwith SIV infected cells.

In certain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing,treating, or ameliorating infections caused by HIV-1 and/or HIV-2. Incertain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing, treatingor ameliorating infections caused by HIV-1 subtype B.

In certain embodiments, the compounds, pharmaceutical compositions, andtherapeutic methods disclosed herein are useful for preventing,treating, or ameliorating infections caused by SIV.

The compounds, pharmaceutical compositions, and therapeutic methodsdescribed herein are useful for preventing, treating, or amelioratinginfections caused by filoviruses, including but not limited to: Marburgvirus, Zaire ebolavirus, Sudan ebolavirus, Cote d'Ivoire ebolavirus,Reston ebolavirus and Bundibugyo ebolavirus.

The compounds, pharmaceutical compositions, and therapeutic methodsdescribed herein are useful for preventing, treating, or amelioratinginfections caused by coronaviruses including, but not limited to,SARS-CoV-2, SARS-CoV and MERS-CoV.

Antiviral Evaluation Using the “One-Stone-Two-Birds” Pseudo-Type Assay.

Production of HIV Pseudovirions. This protocol is designed to identifypotential inhibitors for HIV, coronavirus, Ebola, Marburg and influenzareplication (post entry steps). HIV/VSVG or HIV/SARSP or HIV/HA orHIV/EBOV or HIV/MARV virions were produced, respectively, byco-transfecting with either 0.5 μg VSVG (vesicular stomatitis virusglycoprotein) envelope expression plasmid 0.5 μg SARSP (SARS-CoV-2 spikeprotein) expression plasmid or 0.5 μg hemagglutinin (HA) envelopeexpression plasmid with 0.5 μg neuraminidase (NA) expression plasmid or0.5 μg EBVG (Ebola virus glycoprotein) envelope expression plasmid or0.5 μg MAVG (Marburg virus glycoprotein) envelope expression plasmid and2 μg replication-defective HIV vector (pNL4-3.Luc.RE) into humanembryonic kidney 293T cells (90% confluent) in six-well plates via PEI(polyethylenimine) (Invitrogen, Carlsbad, Calif., USA), as previouslydescribed with a modified procedure. The HIV vector pNL4-3.Luc.RE wasobtained through the AIDS Research and Reference Reagent Program(Division of AIDS, NIAID, NIH). Sixteen hours post-transfection, allmedia were replaced with fresh, complete DMEM. Eight hourspost-transfection, all media were replaced with fresh complete DMEM.Forty-eight hours post-transfection, the supernatants were collected andfiltered through a 0.45-μm-pore-size filter (Millipore, Billerica,Mass., USA) and the pseudo virions were directly used for infection.

Anti-HIV and anti-H5N1 Influenza Virus Evaluation Assay. This protocolis to identify potential inhibitors for HIV and influenza virusreplication (post-entry steps). In this system, the HIV vectorpNL4-3.Luc.RE was co-transfected with the VSVG to generate HIV/VSVGvirions (HIV virion with VSV glycoprotein on the viral surface), and thesame HIV vector was co-transfected with the H5N1 HA and NA constructs togenerate HIV virions with bird flu HA on the viral surface [HIV/HA (HIVvirion with HA and NA glycoproteins on the viral surface)]. This pNL4-3was derived from an infectious molecular clone of a SI (syncytiuminducing), T-tropic virus, which is replication deficient since the HIVis Env⁻ and Vpr⁻. In addition, the luciferase gene (luc) carried by thisrecombinant HIV vector served as the reporter for HIV replication(reverse transcription, integration and HIV gene expression).

The infection level was measured as relative light units (RLUs) in theinfected cells. The luciferase activities of the 293T cells infectedwith the HIV vector pNL4-3.Luc.RE reached the range of 10⁵-10⁶ RLUs,approximately 100-fold higher than the background levels when measuredusing the HIV virions without VSVG. The evaluation principle is that thelevel of the luciferase activity in the cells should be proportional tothe level of viral entry and replication. If a sample can interfere withHIV replication/or HA-mediated viral entry, the level of the luciferaseactivity in the infected cells will be reduced. Thus, using thisprotocol, a sample capable of inhibiting HIV or influenza virusreplication was identified. The test fractions or compounds wereevaluated as follows. Target A549 human lung cells were seeded at0.5×10⁵ cells per well (24-well plate) in complete DMEM. The lung cellline was used since it is susceptible to HA-mediated viral entry. Thestock HIV/VSVG or HIV/HA virions (approximately 2×10⁶ relative lightunits, or RLUs, on the target cells) were mixed with the individualsample first, and the mixture was incubated with the A549 target cellsfor 24 hours. Ten microliters of serial concentrations (for example, 20,10, 5, 2.5, 1.25, 0.625 and 0.3125 μg/mL) and 190 μL of the pseudoviruswere incubated with target cells. Twenty-four (24) hours post-infection,all media containing sample and virus was removed from target cells andreplaced with fresh and complete DMEM. Forty-eight (48) hourspost-infection, the target cells were lysed and the luciferase activitywas determined. Two different outcomes may occur: 1) It is likely thatsome samples will “inhibit replication” of both HIV/VSVG and HIV/HAvirions (lower Luc for HA and VSVG), since some of these samples canblock post-entry steps during viral entry, or some of them are justtoxic to the target cells. These samples are classified as anti-HIV. 2)The samples which can specifically inhibit the HIV/HA viral entry (lowerLuc for HIV/HA, but not for HIV/VSVG) will be classified as anti-HAinhibitors (influenza virus inhibitors). The concentration of druginhibiting 50% of virus infectivity (EC₅₀ value) was determined.

Anti-HIV, anti-Ebola and anti-Marburg Virus Evaluation Assay. Thisprotocol was modified from the aforementioned anti-H5N1 influenza virusevaluation assay, which was designed to identify potential inhibitorsfor HIV, Ebola and Marburg viruses replication (post-entry steps). Inthis system, the HIV vector pNL4-3.Luc. R.E. was co-transfected with theVSVG to generate HIV/VSVG virions, and the same HIV vector wasco-transfected with the Ebola or Marburg glycoprotein (GP) constructs togenerate HIV virions with Ebola or Marburg GP on the viral surface(HIV/EBVG or HIV/MAVG). The infection level was measured as relativelight units (RLUs) in the infected cells. The luciferase activities ofthe 293T cells infected with the HIV vector pNL4-3.Luc. R.E. reached therange of 10⁵-10⁶ RLUs, approximately 100-fold higher than the backgroundlevels when measured using the HIV virions without VSVG. The evaluationprinciple is that the level of the luciferase activity in the cellsshould be proportional to the level of viral entry and replication. If acompound can interfere with HIV replication/or EBVG or MAVG-mediatedviral entry, the level of the luciferase activity in the infected cellswill be reduced. Thus, using this protocol, compounds capable ofinhibiting HIV, EBOV and MARV replication were identified. The testcompounds were evaluated as follows. Target A549 human lung cells wereseeded at 0.5×10⁵ cells per well (24-well plate) in complete DMEM. Thelung cell line was used since it is susceptible to EBVG or MAVG-mediatedviral entry. The stock HIV/VSVG or HIV/EBVG or HIV/MAVG virions(approximately 2×10⁶ relative light units, or RLUs, on the target cells)were mixed with the individual sample first, and the mixture wasincubated with the A549 target cells for 24 hours. Ten microliter ofeach sample in varying concentrations and 190 μL of the pseudovirus wereincubated with target cells. Twenty-four (24) hours post-infection, allmedia containing sample and virus was removed from target cells andreplaced with fresh and complete DMEM. Forty-eight (48) hourspost-infection, the target cells were lysed and the luciferase activitywas determined.

Anti-SARS-CoV-2 Evaluation Assay. This protocol was modified from theaforementioned anti-H5N1 influenza virus evaluation assay, which wasdesigned to identify potential inhibitors for SARS-CoV-2. In this assay,the HIV vector pNL4-3.Luc. R.E. was co-transfected with SARS-CoV-2 spikeprotein (SARSP) expression plasmid to generate SARS-CoV-2 pseudovirions(HIV/SARSP). Target Hep G2 liver cancer cells were seeded at 4×10³ cellsper well (96-well plate) in complete EMEM. The liver cell line was usedbecause it is susceptible to SARS-CoV-2 mediated viral entry. Tenmicroliter of each sample in varying concentrations and 190 μL of theHIV/SARSP were incubated with target cells. Forty-eight (48) hourspost-infection, the target cells were lysed and the luciferase activitywas determined. Arbidol was used as positive control in the experiments.The IC₅₀ value of arbidol against SARS-CoV-2 pseudovirion was measuredas 5.23 μM, which was in agreement with the literature reported value(4.11 μM).

Anti-HIV Evaluation Using HIV-1 Clinical Strains.

The HIV-1 clinical strains such as BAL and SF162 (macrophage-tropic:M-tropic), BAL (T-cell line tropic: T-tropic), and 89.6 (a dual tropicstrain), HIV-1_(LAV) (wild type), NRTI (nucleoside reverse transcriptaseinhibitor)-resistant isolate (HIV-1_(1617.1)) (AZT resistant strain fromAIDS repository) and NNRTI (non-nucleoside reverse transcriptaseinhibitor)-resistant isolate (HIV-1_(N119)) (nevaripine resistant strainfrom AIDS repository) were used in the study. A standardized humanperipheral blood mononuclear cell culture (PBMC) assay was used todetermine the compound susceptibility of these HIV-1 strains. AZT, ananti-HIV drug in clinical use, was used as a positive control. All datawere generated from three independent experiments, each performed intriplicate. Prior to HIV-1 infection, fresh human PBMCs were used ineach experiment. Briefly, donor PBMCs were suspended in R-3 medium [RPMI1640 medium supplemented with 15-20% FBS (fetal bovine serum), 5% IL-2(human interleukin-2), 250 U of penicillin per mL, 250 μg ofstreptomycin per mL and 2 mM L-glutamine] was stimulated with PHA(phytohaemagglutinin, 2-3 μg/mL) for seven days. The preparations(samples) were added to the cultured cells, and the different HIV-1strains were used to challenge the cultured cells in 96-well plates[1×10⁵ cells per well with 1000 TCID₅₀ (virus 50% tissue cultureinfectious doses) of HIV strain]. After seven days of incubation, thesupernatants were collected and the HIV p24 levels of the infected cellswere determined using a p24 antigen ELISA. To measure IC₅₀ values, eachdrug was tested using a serial of concentrations (for example, 5, 1,0.2, 0.04, 0.008, 0.016 and 0 μg/mL). The IC₅₀s were calculated bycomparing p24 antigen values for the samples-containing wells with thosefor no drug control wells. For the p24 assay, the maximum cutoff shouldbe around 120-150 μg/mL.

Antiviral Evaluation Using Infectious Influenza Viruses.

A panel of influenza viruses such as influenza H1N1 (A/HK/415742/09),H3N2 (A/Hong Kong/I/1968), H5N1 (A/Vietnam/1203/2004H), H7N1(A/Rhea/North Carolina/39482/93), H7N7 (A/Netherlands/219/2003), H7N9(A/Anhui/1/2013) and H9N2 (A/Chicken/Y280/97) were used in the studies.Samples were evaluated for their antiviral activities against theinfluenza viruses in A549 cells. Briefly, the preparations (samples)were added to the cultured cells, and the different influenza strainswere used to challenge the cultured cells in 24-well plates (1×10⁵ cellsper well). After removal of the unbound viruses, the cells wereincubated for 48 h. The viral supernatants were collected and viraltiters were determined by standard plaque assay in MDCK (Madin-Darbycanine kidney) cells.

Cytotoxicity Evaluation Using the SRB Assay.

The cytotoxicity of the sample for A549 cells was measured using thesulforhodamine B (SRB) assay (Vichai V, Kirtikara K. Nature protocols2006; 1: 1112-1116). Briefly, 190 μL of A549 cells (2×10⁴ cells/mL) wasseeded in each well of a 96-well cell culture plate. After 24 hours, 10μL of DMSO alone, 10 μL of zidovudine (AZT) as positive controlin 10%DMSO, and 10 μL of each sample in 10% (v/v) DMSO were respectively addedinto wells of a 96-well tissueculture plate. 10 μL of 10% (v/v) DMSO wasadded into each blank well of a 96-well tissueculture plate as abackground calculation plate. After incubation at 37° C. for 2 days, 50μL cold 50% (w/v) trichloroacetic acid (TCA) were added into each wellof the plates, and were further incubated at 4° C. for 1 h. The plateswere then washed four times with low-running tap water, and they wereallowed to dry at room temperature (r.t.). 50 μL of 0.4% (w/v) SRBsolution was added to each well. The plate was left at r.t. for 5˜10mins and were quickly rinsed with 1% (v/v) acetic acid to remove unbounddye. The plates were allowed to dry at r.t. 100 mL of 10 mM Tris basesolution (pH 10.5) was added to each well and the plates were shake on agyratory shaker for at least 30 min to solubilize the protein-bound dye.The OD values were measured at 515 nm in a microplate reader. The CC₅₀(the concentration of an agent causing 50% cytotoxicity) values werecalculated using the GraphPad Prism version 5.0 (GraphPad Software, SanDiego, Calif.).

Toxicity Evaluation in Mice. Repeated-dose toxicity study in mice wasapplied on the selected samples. The animal study was approved andperformed according to Animal Care and Use Guidelines of the AnimalEthics Committee at Hong Kong Baptist University and performed followingAnimal Care and Use guidelines set by NIH (National Institute of Health,USA). BALB/c nude mice, SPF class, male or female, 6-8 weeks old, werepurchased from Charles River Laboratories. Before the experiment, themice are kept for one week of acclimatization to SPF class laboratoryconditions. The mice were then divided into three groups: two dose (25and 50 mg/kg: 10 mice/each dose group) groups of an ANL compound and onedose of vehicle (negative control: 10 mice). Daily injections at i.p.sites were scheduled for 28 days. Weights of mice were measured twice aweek until the end of the experiment. Skin conditions, food intake,water consumption and posture of mice were also inspected. All mice weresacrificed as the end of the experiment to inspect the essential organssuch as liver, heart, kidney, lung and spleen.

TABLE 1 Anti-HIV activity of ANLs.^(a) Compound No. CC₅₀ (nM) EC₅₀ (nM)SI  5 3607 37.1 97.3 (diphyllin) 12a >10111 517.7 >19 12b >26800206.7 >129 13 >114000 23143.0 >4.9 14a 548.5 102.7 5.3 14b 2124.5 277.97.6 15a 918.1 33.6 27.3 15b 1331.1 15.5 85.7 16 3397.3 NE — 17a 45235.172.0 627.9 17b 2143.9 2.6 815.3 17c 889.8 4.8 186.9 17d 61056.9 1588.638.4 17f 7073.4 772.5 9.2 17g 1322.0 79.9 16.6 17h 1457.9 96.5 15.1 181360.9 487.6 2.8 19a 10494.5 77.2 136.0 19b 77.3 6.1 12.7 19c 1672.988.9 18.8 19d 5426.1 22.9 237.4 19e >9551 211.7 >45 19f 4505.2 975.9 4.619g 196.2 19.3 10.2 19h 2392.3 270.0 8.9 19i 38358.6 877.1 43.7 19k1064.9 60.1 17.7 19l 4185.5 75.9 55.1 19m 1496.5 88.6 16.9 19n 1056.339.5 26.8 19o 1218.8 35.7 34.2 19p >9198 249.3 >36 20 388.8 21.7 17.9 21207.2 17.8 11.7 22 358.23 1.61 222.2 23 1581 18.8 84 24 >11864 88.9 >13325a 3083.4 7.0 438.6 25b 259.1 NE — 26a 6220.8 NE — 26b 347.0 21.1 16.4(patentiflorin A) 26c >9756.7 NE — 26d >9496.9 42.3 >426.1 26e >9496.9NE — 26f >9216.8 NE — 26g 7767.9 1402.8 5.5 27ab 2757.4 NE — 27ac 2936.0NE — 27ad 782.7 NE — 27ae 2593.3 NE — 27af 2251.3 NE — 27ba 1162.3 8.47137.3 27bb 589.2 1.01 584.1 27bc 1365.1 0.93 1472.4 27bd 420.8 6.39 65.927be 1668.9 1.96 852 27bf 308.2 1.17 263.9 28ab1 512.7 NE — 28ab2 386.02.3 167.7 28ab3 385.5 22.9 16.8 29a 438.3 NE — 29b 187.2 4.7 40.2 30a145.3 NE — 31a 55.8 NE — 31b 684.0 13.4 51.0 31c 13.4 0.03 477.0 31d >631.36 >74 31e 17.7 2.41 7.3 31f 3.74 NE — 31g 26.7 NE — 31h 336.5 NE —311 5733.7 14.4 399.6 31j 6.68 0.91 7.3 31k 507.8 NE — 32a 52.7 NE — 32b52.7 NE — 32c 52.7 NE — 32d 52.7 NE — AZT^(c) 2.6 ^(a)Results areexpressed as CC₅₀ (the concentration caused inhibition of cell growth ofhost A549 cells by 50%) and EC₅₀ (effective concentration of compound toinhibit viral growth by 50%) values in nM, and data were obtained fromtriplicate experiments. SI = CC₅₀/EC₅₀ b NE: No effect at the CC₅₀tested in the host A549 cells ^(c)Positive control compound.

TABLE 2 Anti-H5N1 viral activity of ANLs.^(a) % Inhibition CompoundConcentration against No. (ng/mL) H5N1 virus 26a 5000 97.8% 1000 93.6%26e 5000 88.9% 1000 48.5% 27ab 5000 97.8% 1000 96.9% 29a 250 97.3% 5059.4% 30a 250 97.6% 50 95.7% 12.5 44.1% 250 97.6% 31a 50 94.4% 12.587.9% 31e 250 98.4% 50 97.9% 12.5 98.1% 31g 250 98.1% 50 97.9% 12.597.9% 31h 250 97.6% 50 94.6% 31j 250 97.9% 50 87.6% 31k 250 95.7% 5086.0% 31l 250 96.8% 50 96.0% 12.5 82.8% AZT^(b) 20 96.5% ^(a)Results arefrom three triplicate experiments. ^(b)Positive control compound.

TABLE 3 Inhibitory effects of selected ANLs against H5N1 virus, EBOV,VSV and A549 cells.^(a) % % Inhibition % % Inhibition Concen- againstInhibition Inhibition against Compound tration H5N1 against against A549No. (ng/mL) virus EBOV VSV cells 19p 5000 96.5 89.4 — 17.2 1000 82.158.1 — −13.0 26f 500 87.2 89.1 70.8 18.2 100 62.4 82.7 47.18  −3.8428ab1 50 86.0 82.4 83.5 40.9 12.5 91.8 81.0 85.9 35.5 31a 5000 98.4 95.299.6 99.8 1000 94.7 81.8 80.4 47.5 31e 50 96.5 86.5 86.3 66.8 12.5 95.477.5 77.4 53.1 31g 50 89.0 90.9 95.7 54.6 10 78.9 79.1 63.3 11.0 31j 5098.2 96.6 99.9 93.8 12.5 97.5 93.9 98.4 77.0 31l 50 95.6 94.2 98.4 82.112.5 95.1 86.4 88.7 52.0 32b 50 88.1 92.7 99.6 76.0 10 91.7 86.4 90.940.5 32c 500 88.1 88.2 88.1 39.7 100 65.1 85.5  48.87 2.9 32d 500 83.582.7 71.7 24.3 100 19.3 65.5 100.2  −2.0 AZT^(b) 20 92.7 93.6 99.4 1.1Toremifene^(b) 500 28.4 93.6  6.58 −15.1 ^(a)Results are from threetriplicate experiments. ^(b)Positive control compounds.

Compound 26b (patentiflorin A) was identified as an anti-HIV leadcompound from both methanol extracts of J. cf. patentiflora (the rootsand stems of) and J. procumbens (aerial parts), and compound 5(diphyllin) was isolated as a pure compound from the methanol extract ofJ. procumbens (aerial parts). Compound 5 was also obtained as anintermediate through the total synthesis of 26b in our previous studies.Compounds 26b and 5 showed anti-HIV activities with EC₅₀ values of 21.1and 37.1 nM and SIs of 16.4 and 97.3, respectively, in our present“One-Stone-Two-Birds” evaluation system. In order to improve theirantiviral activities and reduce their cytotoxicities, we synthesizednumerous analogues of the two compounds. We designed synthetic route toreplace the hydroxyl group of 5 with different functional groups, whichled to the preparation of compounds 12-24 (FIG. 2 ). We further designedsynthetic route to replace the 6-deoxymethyl glucose sugar unit of 26bwith different sugar units or to selectively substitute one or two orthree or four hydroxyl groups on the sugar moieties of 26a-g, whichresulted in the preparation of compounds 25-32 (FIGS. 4-6 ). Thesecompounds were evaluated for their antiviral activities in the“One-Stone-Two-Birds” system. As shown in the results of Table 1,compounds 17a, 17b, 17c, 19a, 19d, 22, 25a, 26d, 27bb, 31c and 31i,displayed much more improved anti-HIV activity than patentiflorin A(26b) and diphyllin (5) in terms of SI values. It was observed that incertain instances the stereochemistry of the sugar units is essentialfor the biological activities among these synthesized diphyllinglycosides. Compared with 26e, the anti-HIV activity of 26d was muchhigher (26d, EC₅₀=42.3 nM, and SI>426.1; 26e, no inhibition at 9496.9nM). The high anti-HIV activity and selectivity of 26d were achieved byreplacing the D-(+)-fucose in 26e with L-(−)-fucose in 26d. Compounds12b, 17a, 19a and 21 were selected to be further evaluated against abroad spectrum of HIV-1 clinical and resistant strains. Compound 12bshowed inhibitory effects against SF162, 89.6, Bal, Lav.04, 1617-1 andN119 with IC₅₀ values of 340, 376, 458, 312, 537 and 638 nM,respectively (the CC₅₀ value against PBMC at >100 μM). Compound 17ashowed inhibitory effects against SF162, 89.6, Bal, Lav.04, 1617-1 andN119 with IC₅₀ values of 122, 134, 168, 151, 174 and 203 nM,respectively (the CC₅₀ value against PBMC at >100 μM). Compound 19ashowed inhibitory effects against SF162, 89.6, Bal, Lav.04, 1617-1 andN119 with IC₅₀ values of 154, 143, 188, 138, 152 and 185 nM,respectively (the CC₅₀ value against PBMC at >100 μM). Compound 21showed inhibitory effects against SF162, 89.6, Bal, Lav.04, 1617-1 andN119 with IC₅₀ values of 23.9, 28.7, 26.5, 30.1, 39.3 and 42.1 nM,respectively (the CC₅₀ value against PBMC at 25.6 μM). Compounds 19p,26f, 28ab1, 31a, 31e, 31g, 31j, 31l, 32b, 32c and 32d were selected tobe tested for their inhibitory effects in several concentrations againstH5N1 virus, EBOV (Ebola virus) and VSV (vesicular stomatitis virus) incomparison with their inhibitory effects against the A549 host cells(Table 3). Compounds 28ab1, 31e, 31g, 31l, 31j and 32b showed more than77% inhibitory effects against the three viruses (H5N1 virus, EBOV andVSV) in a concentration of below 12.5 ng/mL. The compound series of26-32 were repeatedly tested for their antiviral activities against H5N1virus, EBOV and VSV. Compounds 26b, 26f, 26g, 28ab1, 28ab2, 28ab3, 29a,30a, 31e, 31j, 31l, 32a, 32b, 32c, 32d and AZT (the positive control)were measured to show inhibitory effects against H5N1 virus with EC₅₀values of 4.48, 371.3, 63.6, 5.05, 0.23, 1.16, 5.57, 3.88, 0.74, 0.84,0.013, 0.63, 0.77, 5.20, 199.8 and 3.48 nM, respectively. Compounds 26b,26f, 26g, 28ab1, 28ab2, 28ab3, 29a, 30a, 31e, 31j, 31l, 32a, 32b, 32c,32d, AZT (the positive control) and toremifene (the positive control)were measured to show inhibitory effects against EBOV with EC₅₀ valuesof 54.8, 515.6, 123.0, 18.9, 16.3, 0.98, 10.6, 1.84, 1.13, 0.84, 0.19,1.20, 0.85, 20.3, 395.7, 3.80 and 212.1 nM, respectively. Compounds 26b,26f, 26g, 28ab1, 28ab2, 28ab3, 29a, 30a, 31e, 31j, 31l, 32a, 32b, 32c,32d and AZT (the positive control) were measured to show inhibitoryeffects against VSV with EC₅₀ values of 42.5, 1035, 210.1, 3.78, 4.98,0.85, 10.6, 8.60, 2.41, 0.91, 0.37, 18.1, 3.15, 123.9, 286.1 and 7.06nM, respectively. Compounds 5, 26b, 26e, 28ab1, 31j, 32a, 32b, 32c, 32dand arbidol (the positive control) were measured to show inhibitoryeffects against SARS-CoV-2 with EC₅₀ values of 150.1, 67.2, 30.4, 26.3,10.3, 2.95, 3.01, 50.8, 80.4, and 4,110 nM, respectively. Compounds 26b,26f, 26g, 28ab1, 28ab2, 28ab3, 29a, 30a, 31e, 31j, 31l, 32a, 32b, 32cand 32d and taxol (the positive control) were measured to showinhibitory effects against the host cell line A549 with CC₅₀ values of386.1, 8649, 4129, 190.0, 108.2, 6.14, 121.6, 35.2, 17.7, 6.68, 4.85,61.6, 30.7, 1038, 1349 and 2.99 nM, respectively. Compounds 17a and 25awere further evaluated for its toxicity in mice with each having twodoses at 25 and 50 mg/kg in comparison with a vehicle control group. Atthe end of the animal study, no toxic symptoms (weight loss, skindamage, food and water intake impairment and postual abnormality) andmortality were observed for the mice of all groups. The excised organsof all mice appeared as normal at the end of the experiment.

Materials and Methods

General Experimental Procedures

1D and 2D NMR spectra were recorded on a Bruker DRX-400 MHz spectrometer(Rheinstetten, Germany). Chemical shifts (δ) were expressed in ppm andcoupling constants (J) are reported in Hz. All NMR experiments wereobtained by using standard pulse sequences supplied by the vendor.Chemical shifts (δ) were expressed in ppm with reference to the solventsignals (CD₃OD: ¹H: 3.31 ppm, ¹³C: 49.00 ppm; CDCl₃: ¹H: 7.27 ppm, ¹³C:77.23 ppm; DMSO-d₆: ¹H: 2.50 ppm, ¹³C: 39.51 ppm; acetone-d₆: ¹H: 2.05ppm, ¹³C: 29.92, 206.68 ppm.), and coupling constants (J) are reportedin Hz. Column chromatography was carried out on silica gel (230-400mesh, Natland International Corporation, Research Triangle Park, N.C.,USA). Thin-layer chromatography (TLC) was performed on EMD glass-backedplates coated with 0.25 mm layers of silica gel 60 F254 (Kassel,Germany). HRTOFMS spectra were recorded on a Micromass QTOF-2 (Milford,Mass., USA), an Agilent 6540 Q-TOF (Santa Clara, Calif., USA), anAgilent 6460 Triple Quadrupole, or a Bruker Q-TOF mass spectrometer(Bremen, Germany). All reagents were purchased from commercial sourcesand used without further purification.

Compound 5

To a suspension of the crude 2-bromo-4,5-dimethoxybenzaldehyde (30 g,125 mmol) in toluene (1 L) was added ethylene glycol (14 mL, 250 mmol)and p-toluenesulfonic acid (TsOH) (5.2 g, 25 mmol). A Dean-Starkapparatus filled with toluene (PhMe) was fitted to the round bottomflask, and the reaction was refluxed overnight. After cooling to roomtemperature (r.t.), toluene was removed under reduced pressure. Thecrude mixture was then dissolved in ethyl acetate (EtOAc) and washedwith sodium bicarbonate (NaHCO₃) aqueous, water, then brine, dried withanhydrous sodium sulfate (Na₂SO₄), and concentrated under reducedpressure. Separation of the mixture on silica gel flash chromatography(deactivated silica gel, 16%-25% EtOAc in n-hexane) afforded2-(2-bromo-4,5-dimethoxyphenyl)-1,3-dioxolane (2) as an off-white solid(33.5 g, 95%). To a stirring solution of the acetal (2) (30 g, 104 mmol)in 1 L of tetrahydrofuran (THF), after cooled to −78° C., n-buyllithium(n-BuLi) (1.8 M, 69.3 mL, 124.8 mmol) was added drop-wise for anadditional 30 min. Following the addition, the reaction mixture wasstirred for 30 min at −50° C., and then piperonal (114.4 mmol) wasadded. The mixture was stirred at −78° C. for 0.5 h, and then thereaction was allowed to warm to r.t. for an additional 2 h. The reactionwas then quenched with water, and the aqueous layer was extracted withEtOAc. The combined organic layers were washed with water, then brine,dried with Na₂SO₄, and concentrated under reduced pressure to afford apale-yellow syrup (3), which was immediately subjected to a Diels-Alderreaction. To a solution of the crude intermediate in dichloromethane(CH₂Cl₂) (100 mL) was added acetic acid (AcOH) (35 mL) and dimethylacetylenedicarboxylate (DMADC) (16.7 mL, 104 mmol). The reaction wasstirred at 140° C. for 12 h. Upon cooling to r.t., the reaction wasdiluted with water, and the aqueous layer was extracted with CH₂Cl₂. Thecombined organic layers were washed, dried and filtered through a plugof silica gel to remove the majority of impurities. The resulted redviscous oil was purified by recrystallization in ethanol (EtOH). Thecrude diester (4a, 5 mmol) was dissolved in THF (70 mL) with thedrop-wise addition of sodium borohydride (NaBH₄) (0.945 g, 25 mmol), andthe reaction was stirred at 80° C. for 12 h. The reaction was thencooled back to r.t. The resulting slurry was acidified to a pH value ofapproximately 2 with dilute HCl (2 M). Following acidification, theaqueous layer was extracted with EtOAc, and the combined organic layerwas washed with water, then brine, dried with Na₂SO₄ and concentrated.The resulted red viscous oil was purified by silica gel columnchromatography to give the target compound 5. HR-EIMS: m/z [M+H]⁺381.0974 (calcd. 381.0912). ¹H NMR (400 MHz, acetone-d₆) δ 7.69 (11H, s,H-2), 7.09 (1H, s, H-5), 6.96 (11H, d, J=7.9 Hz, H-5′), 6.85 (1H, d,J=1.6 Hz, H-2′), 6.91 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.09 (1H, br s,OCH₂O), 6.07 (1H, br s, OCH₂O), 5.37 (2H, s, H-9), 3.99 (3H, s, 3-OCH₃),3.73 (3H, s, 4-OCH₃).

Compound 12a

To a solution of diphyllin (5, 190 mg, 0.5 mmol) and t-BuOCOCH2Br (193mg, 1 mmol) in DMF (5 mL) was added Cs₂CO₃ (326 mg, 1 mmol). The mixturewas stirred at r.t. for 2 h. After quenched with water, the aqueouslayer was then extracted with EtOAc and the combined organic layer werewashed with water, then brine, dried with Na₂SO₄ and concentrated underreduced pressure. The mixture was purified by prep-TLC to afford alight-yellow solid (12a, 247 mg, 99%). HR-EIMS: m/z [M+H]⁺ 495.1638(calcd. 495.1655). ¹H NMR (400 MHz, CDCl₃) δ 7.73 (1H, s, H-2), 7.07(1H, s, H-5), 6.96 (11H, d, J=7.9 Hz, H-5′), 6.82 (11H, d, J=1.3 Hz,H-2′), 6.79 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.09 (11H, br s, —OCH₂O—),6.05 (1H, br s, —OCH₂O—), 5.48 (2H, s, H-9), 4.69 (2H, s, OCH₂C═O), 4.09(3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃), 1.51 (9H, s, t-Bu). ¹³C NMR (100MHz, CDCl₃) δ 169.5 (C-9′), 167.8 (O—C═O), 151.8 (C-3), 150.4 (C-4),147.5 (C-4′, C-7), 146.7 (C-3′), 135.5 (C-7′), 130.8 (C-6), 128.3(C-1′), 126.4 (C-8′), 126.2 (C-1), 123.6 (C-6′), 119.1 (C-8), 110.7(C-2′), 108.2 (C-5′), 106.2 (C-5), 101.3 (C-2), 100.9 (—OCH₂O—), 82.9(t-Bu), 69.8 (OCH₂C═O), 66.4 (C-9), 56.2 (3-OCH₃), 55.9 (4-OCH₃), 28.1(t-Bu).

Compound 12b

To a solution of diphyllin (5, 38 mg, 0.1 mmol) and EtOCOCH₂Br (33 mg,0.2 mmol) in 1 mL DMF was added Cs₂CO₃ (65 mg, 0.2 mmol). The mixturewas stirred at r.t. for 2 h. After quenched with water, the aqueouslayer was then extracted with EtOAc and the combined organic layers werewashed with water, then brine, dried with Na₂SO₄ and concentrated underreduced pressure. The mixture was purified by prep-TLC to afford 12b (46mg, 99%). HR-EIMS: m/z 467.1316 [M+H]+(calcd. 467.1342). ¹H NMR (400MHz, DMSO-d₆) δ 7.73 (1H, s, H-2), 7.03 (1H, d, J=7.9 Hz, H-5′), 6.96(1H, s, H-5), 6.87 (1H, d, J=1.5 Hz, H-2′), 6.75 (1H, dd, J=7.9, 1.7 Hz,H-6′), 6.12 (2H, br s, —OCH₂O—), 5.59 (2H, s, H-9), 5.02 (2H, s,OCH₂C═O), 4.23 (2H, q, J=7.1 Hz, Et), 3.95 (3H, s, 3-OCH₃), 3.66 (3H, s,4-OCH₃), 1.24 (3H, t, J=7.1 Hz, Et). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.0(C-9′), 169.0 (O—C═O), 151.4 (C-3), 150.1 (C-4), 147.0 (C-3′), 146.9(C-4′), 146.0 (C-7), 133.8 (C-7′), 129.6 (C-6), 128.2 (C-1′), 126.0(C-8′), 125.6 (C-1), 123.6 (C-6′), 118.8 (C-8), 110.8 (C-2′), 108.0(C-5′), 105.5 (C-5), 101.2 (C-2), 100.9 (—OCH₂O—), 68.7 (OCH₂C═O), 66.4(C-9), 61.0 (Et), 55.7 (3-OCH₃), 55.3 (4-OCH₃), 14.1 (Et).

Compound 13

To a solution of 12a (247 mg, 0.5 mmol) in 10 mL CH₂Cl₂, 1 mL DMF and 2mL TFA was added. The mixture was stirred for 12 h at r.t. TLC showedthat the reaction had completed and a new spot with higher polarityappeared. After the reaction mixture was concentrated, EtOH was added topromote the precipitation. The solid was collected to afford 13 (210 mg,95%). HR-EIMS: m/z [M+H]⁺ 439.100 (calcd. 439.1029). ¹H NMR (400 MHz,DMSO-d₆) δ 7.78 (1H, s, H-2), 7.04 (1H, d, J=8.0 Hz, H-5′), 6.97 (1H, s,H-5), 6.89 (1H, d, J=1.5 Hz, H-2′), 6.77 (11H, dd, J=7.9, 1.7 Hz, H-6′),6.13 (2H, s, —OCH₂O—), 5.76 (1H, s, OH), 5.60 (2H, s, H-9), 4.93 (2H, s,OCH₂C═O), 3.96 (3H, s, 3-OCH₃), 3.67 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz,DMSO-d₆) δ 170.9 (C-9′), 169.4 (O—C═O), 151.7 (C-3), 150.5 (C-4), 147.4(C-3′), 147.3 (C-4′), 146.7 (C-7), 134.1 (C-7′), 130.0 (C-6), 128.7(C-1′), 126.3 (C-6′), 126.1 (C-8′), 124.0 (C-1), 119.3 (C-8), 111.3(C-2′), 108.4 (C-5′), 105.9 (C-5), 101.6 (C-2), 101.5 (—OCH₂O—), 69.1(OCH₂C═O), 66.8 (C-9), 56.1 (3-OCH₃), 55.7 (4-OCH₃).

Compound 14a

To a solution of 13 (44 mg, 0.1 mmol) in CH₂Cl₂ (10.00 mL) were addedEt₃N (30 mg, 0.3 mmol) NH₄Cl (16 mg, 0.3 mmol) and HATU (114 mg, 0.3mmol) at 25° C., the reaction was stirred at 25° C. for 16 h. Afterquenched with water, the aqueous layer was then extracted with EtOAc andthe combined organic layer were washed with water, then brine, driedwith Na₂SO₄ and concentrated under reduced pressure. The mixture waspurified by prep-TLC to afford 14a (40 mg, 92%). HR-EIMS: m/z 438.1167[M+H]+(calcd. 438.1189). ¹H NMR (400 MHz, DMSO-d₆) δ 7.80 (11H, s, NH₂),7.70 (11H, s, H-2), 7.55 (1H, s, NH₂), 7.04 (1H, d, J=7.9 Hz, H-5′),6.97 (1H, s, H-5), 6.89 (1H, d, J=1.5 Hz, H-2′), 6.77 (1H, dd, J=7.9,1.6 Hz, H-6′), 6.12 (2H, s, —OCH₂O—), 5.60 (2H, s, H-9), 4.70 (2H, s,OCH₂C═O), 3.97 (3H, s, 3-OCH₃), 3.66 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz,DMSO-d₆) δ 170.1 (C-9′), 169.0 (O—C═O), 151.3 (C-3), 150.0 (C-4), 147.0(C-3′), 146.9 (C-4′), 146.1 (C-7), 133.7 (C-7′), 129.6 (C-6), 128.3(C-1′), 126.1 (C-8′), 125.7 (C-1), 123.6 (C-6′), 118.9 (C-8), 110.9(C-2′), 108.0 (C-5′), 105.5 (C-5), 101.2 (C-2, —OCH₂O—), 70.6 (OCH₂C═O),66.5 (C-9), 55.8 (3-OCH₃), 55.3 (4-OCH₃).

Compound 14b

To a solution of 13 (44 mg, 0.1 mmol in CH₂Cl₂ (10.00 mL) and DMF (1 mL)were added Et₃N (30 mg, 0.3 mmol) and MeNH₂ (0.3 mL, 0.3 mmol) and HATU(114 mg, 0.3 mmol) at 25° C., the resulting mixture was stirred at 25°C. for 16 h. After quenched with water, the aqueous layer was thenextracted with EtOAc and the combined organic layer were washed withwater, then brine, dried with Na₂SO₄ and concentrated under reducedpressure. The mixture was purified by prep-TLC to afford 14b (30 mg,66%). HR-EIMS: m/z 452.1312 [M+H]+(calcd. 452.1345). ¹H NMR (400 MHz,DMSO-d₆) δ 7.61 (1H, s, H-2), 6.96 (1H, d, J=7.9 Hz, H-5′), 6.90 (1H, s,H-5), 6.81 (1H, d, J=1.5 Hz, H-2′), 6.69 (1H, dd, J=7.9, 1.7 Hz, H-6′),6.04 (2H, s, —OCH₂O—), 5.50 (2H, s, H-9), 4.65 (2H, s, OCH₂C═O), 3.90(3H, s, 3-OCH₃), 3.58 (3H, s, 4-OCH₃), 2.60 (3H, s, NH-Me). ¹³C NMR (100MHz, DMSO-d₆) δ 169.0 (C-9′), 168.2 (O—C═O), 151.4 (C-3), 150.1 (C-4),147.0 (C-3′), 146.9 (C-4′), 146.1 (C-7), 133.9 (C-7′), 129.6 (C-6),128.3 (C-1′), 126.5 (C-8′), 125.8 (C-1), 123.6 (C-6′), 118.9 (C-8),110.9 (C-2′), 108.0 (C-5′), 105.5 (C-5), 101.2 (C-2), 101.1 (—OCH₂O—),71.0 (OCH₂C═O), 66.4 (C-9), 55.8 (3-OCH₃), 55.3 (4-OCH₃), 25.6 (NH-Me).

Compound 15a

A mixture of TBAB (16 mg, 0.05 mmol), PdCl₂ (1.7 mg, 0.01 mmol) andK₂CO₃ (6.9 mg, 0.05 mmol) was dissolved in deionized H₂O (2.00 mL) andstirred at 60° C. for 15 mins. Then diphyllin (5, 38 mg, 0.1 mmol) and0.3 mL ethylene oxide were added and stirred at 60° C. for 5 h. TLCshowed the reaction completed and the one main spot with less polarityformed. The mixture was concentrated to give a yellow green solid whichwas washed by EtOH to give desired product 15a (40 mg, 95%). HR-EIMS:425.1207 m/z [M+H]⁺ (calcd. 425.1236).

¹H NMR (400 MHz, DMSO-d₆) δ 7.76 (1H, s, H-2), 7.03 (1H, d, J=7.9 Hz,H-5′), 6.97 (1H, s, H-5), 6.89 (1H, d, J=1.5 Hz, H-2′), 6.77 (1H, dd,J=7.9, 1.6 Hz, H-6′), 6.12 (2H, s, —OCH₂O—), 5.75 (1H, s, OH), 5.58 (2H,s, H-9), 4.28-4.22 [2H, m, O(CH₂)₂OH], 3.95 (3H, s, 3-OCH₃), 3.81 [2H,m, O(CH₂)₂OH], 3.66 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.2(C-9′), 151.2 (C-3), 150.0 (C-4), 146.9 (C-3′), 146.8 (C-4′), 146.6(C-7), 133.3 (C-7′), 129.6 (C-6), 128.4 (C-1′), 126.6 (C-8′), 126.1(C-1), 123.7 (C-6′), 118.9 (C-8), 110.9 (C-2′), 108.0 (C-5′), 105.5(C-5), 101.1 (C-2, —OCH₂O—), 74.4 [O(CH₂)₂OH], 66.6 (C-9), 60.4[O(CH₂)₂OH], 55.7 (3-OCH₃), 55.2 (4-OCH₃).

Compound 15b

Cs₂CO₃ (100 mg, 0.3 mmol) was added to a mixture of 5 (38 mg, 0.1 mmol)and isobutylene oxide (71 mg, 1 mmol) in 2 mL DMF and stirred at 120° C.for 12 h. TLC showed that the reaction had completed with one main spotwith less polarity formed. The mixture was concentrated to give a yellowgreen solid, which was washed by EtOH to give desired product 15b (38mg, 84%). HR-EIMS: m/z [M+H]⁺ 453.1524 (calcd. 453.1549). ¹H NMR (400MHz, DMSO-d₆) δ 7.71 (1H, s, H-2), 7.04 (1H, d, J=7.9 Hz, H-5′), 6.97(1H, s, H-5), 6.89 (1H, d, J=1.6 Hz, H-2′), 6.77 (1H, dd, J=7.9, 1.7 Hz,H-6′), 6.13 (2H, s, —OCH₂O—), 5.63 (2H, s, H-9), 4.03 (2H, s, CH₂), 3.95(3H, s, 3-OCH₃), 3.67 (3H, s, 4-OCH₃), 1.34 (6H, s, Me). ¹³C NMR (100MHz, DMSO-d₆) δ 169.1 (C-9′), 151.1 (C-3), 149.9 (C-4), 146.9 (C-3′),146.8 (C-4′), 146.7 (C-7), 132.8 (C-7′), 129.5 (C-6), 128.4 (C-1′),125.5 (C-8′), 125.5 (C-1), 123.6 (C-6′), 118.9 (C-8), 110.8 (C-2′),107.9 (C-5′), 105.5 (C-5), 101.1 (C-2), 101.0 (—OCH₂O—), 79.9 (CH₂),69.1 (C), 66.6 (C-9), 55.5 (3-OCH₃), 55.2 (4-OCH₃), 26.4 (Me).

Compound 16

A solution of 5 (19 mg, 0.05 mmol) in dry pyridine (1 mL) was added Ac₂O(7.1 μL, 0.075 mmol). The resulting mixture was stirred under r.t.overnight. After the reaction was complete, it was diluted by CH₂Cl₂ andwashed with 10% HCl solution. The combined organic phase was furtherextracted with a saturate aqueous solution of NaHCO₃ and dried overNa₂SO₄. After concentration, the crude product was purified prep-TLC(1:1 n-hexane:ethyl acetate) to give 16 (20 mg, 94.6%) as a colorlesssolid. HR-EIMS: 423.1077 m/z [M+H]+(calcd. 423.1080). ¹H NMR (400 MHz,CDCl₃) δ 7.17 (1H, s, H-2), 7.12 (1H, s, H-5), 6.97 (1H, d, J=7.8 Hz,H-5′), 6.86 (1H, d, J=1.4 Hz, H-2′), 6.83 (1H, dd, J=7.9, 1.7 Hz, H-6′),6.10 (1H, d, J=1.5 Hz, —OCH₂O—), 6.05 (1H, d, J=1.5 Hz, —OCH₂O—), 5.26(2H, s, H-9), 4.06 (3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃), 2.53 (3H, s,OAc).

Compound 17a

To a solution of 5 (38 mg, 0.1 mmol) and bromoacetonitrile (24 mg, 0.2mmol) in 1 mL acetone was added Cs₂CO₃ (65 mg, 0.2 mmol). The mixturewas stirred at r.t. for 2 hours. TLC show one spot with less polarity.The mixture was purified by prep-TLC to give desired product 17a (40 mg,95%). HR-EIMS: m/z [M+H]⁺ 420.1053 (calcd. 420.1083). ¹H NMR (400 MHz,DMSO-d₆) δ 7.52 (1H, s, H-2), 7.05 (1H, d, J=7.9 Hz, H-5′), 7.01 (1H, s,H-5), 6.91 (1H, d, J=1.6 Hz, H-2′), 6.79 (1H, dd, J=1.7, 7.9 Hz, H-6′),6.13 (2H, s, —OCH₂O—), 5.60 (2H, s, CH₂), 5.34 (2H, s, H-9), 3.99 (3H,s, 3-OCH₃), 3.68 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 168.8(C-9′), 151.8 (C-3), 150.2 (C-4), 147.1 (C-3′), 147.0 (C-4′), 144.7(C-7), 135.5 (C-7′), 129.8 (C-6), 128.4 (C-1′), 127.9 (C-8′), 126.0(C-1), 123.6 (C-6′), 118.8 (C-8), 117.1 (C≡N), 110.8 (C-2′), 108.1(C-5′), 105.8 (C-5), 101.2 (C-2), 100.3 (—OCH₂O—), 66.1 (C-9), 58.0(CH₂), 55.9 (3-OCH₃), 55.3 (4-OCH₃).

Compound 17b

To a solution of 5 (19 mg, 0.05 mmmol) in acetone (0.6 mL) was addedCs₂CO₃ (32.5 mg, 0.1 mmol) and the solution was cooled to 0° C.Iodomethane (10.6 mg, 0.075 mmol) was added dropwise and the mixture waswarmed to r.t. The reaction was complete after 12 h as shown by TLC. Themixture was quenched with saturated NH₄Cl solution, extracted withEtOAc, and washed with brine. After drying (Na₂SO₄) the solvent wasremoved under reduced pressure and the residue was purified by prep-TLC(n-hexane:ethyl acetate, 3:1) to afford the product 17b as white solid(17.7 mg, 90%). HR-EIMS: m/z [M+H]⁺ 395.1125 (calcd. 395.1131). ¹H NMR(400 MHz, CDCl₃) (7.54 (1H, s, H-2), 7.05 (1H, s, H-5), 6.95 (1H, d,J=7.8 Hz, H-5′), 6.78 (2H, m, H-2′, H-6′), 6.09 (1H, d, J=1.5 Hz,—OCH₂O—), 6.04 (1H, d, J=1.5 Hz, —OCH₂O—), 5.54 (2H, s, H-9), 4.13 (3H,s, OCH₃), 4.07 (3H, s, 3-OCH₃), 3.80 (3H, s, 4-OCH₃).

Compound 17c

To a solution of 5 (38 mg, 0.1 mmol) and propynyl bromide (30 mg, 0.2mmol) in acetone was added Cs₂CO₃ (65 mg, 0.2 mmol). The mixture wasstirred at r.t. for 2 h. TLC showed one spot with less polarity. Themixture was purified by a silica gel column to give desired product 17c(40 mg, 95%). HR-EIMS: m/z [M+H]⁺ 419.1112 (calcd. 419.1131). ¹H NMR(400 MHz, DMSO-d₆) δ 7.57 (1H, s, H-2), 7.04 (1H, d, J=7.9 Hz, H-5′),6.98 (1H, s, H-5), 6.91 (1H, d, J=1.5 Hz, H-2′), 6.78 (1H, dd, J=7.9,1.6 Hz, H-6′), 6.12 (2H, s, —OCH₂O—), 5.62 (2H, s, H-9), 5.04 (2H, d,J=2.4 Hz, CH₂), 3.96 (3H, s, 3-OCH₃), 3.71 (1H, t, J=2.4 Hz, C≡CH), 3.66(3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.0 (C-9′), 151.4 (C-3),150.1 (C-4), 147.0 (C-3′), 147.0 (C-4′), 145.6 (C-7), 134.2 (C-7′),129.7 (C-6), 128.2 (C-1′), 127.2 (C-1), 126.3 (C-8′), 123.7 (C-6′),118.8 (C-8), 110.9 (C-2′), 108.1 (C-5′), 105.6 (C-5), 101.2 (C-2), 100.8(—OCH₂O—), 79.7 (CH₂), 79.5 (C≡CH), 66.6 (C-9), 55.7 (3-OCH₃), 55.3(4-OCH₃).

Compound 17d

To a solution of 5 (38 mg, 0.1 mmol) and 2-chloropyrimidine (23 mg, 0.2mmol) in 1 mL DMF was added K₂CO₃ (40 mg, 0.3 mmol). The mixture wasstirred for 12 h at 100° C. TLC showed one spot with less polarity. Themixture was purified by prep-TLC to give desired product 17d (35 mg,76%). HR-EIMS: m/z [M+H]⁺ 459.1161 (calcd. 459.1192).1H NMR (400 MHz,DMSO-d₆) δ 8.71 (2H, d, J=4.8 Hz, pyrimidine), 7.37 (1H, t, J=4.8 Hz,pyrimidine), 7.19 (1H, s, 1H-2), 7.08 (1H, s, H-5), 7.08 (1H, d, J=7.9Hz, H-5′), 7.01 (1H, d, J=1.5 Hz, H-2′), 6.87 (1H, dd, J=7.9, 1.7 Hz,H-6′), 6.15 (2H, br s, —OCH₂O—), 5.24 (2H, s, H-9), 3.81 (3H, s,3-OCH₃), 3.69 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, CDCl₃) δ 169.8(pyrimidine), 169.1 (C-9′), 160.1 (pyrimidine), 160.0 (pyrimidine),151.6 (C-3), 150.2 (C-4), 147.1 (C-4′), 147.1 (C-3′), 146.8 (C-7), 134.9(C-7′), 129.9 (C-6), 128.8 (C-1′), 127.7 (C-1), 127.0 (C-8′), 123.8(pyrimidine), 123.8 (C-6′), 119.2 (C-8), 110.9 (C-2′), 108.4 (C-5′),106.2 (C-5), 101.0 (C-2), 100.8 (—OCH₂O—), 66.7 (C-9), 56.03 (3-OCH₃),55.4 (4-OCH₃).

Compound 17e

5 (38 mg, 0.1 mmol), Et₃N (30 mg, 0.3 mmol) and DMAP (catalyzed amount)were dissolved in 2 mL of dry CH₂Cl₂. Cyclopropanecarbonyl chloride (31mg, 0.3 mmol) was added to the mixture at 0° C. The reaction mixture wasallowed to warm up to 25° C. and kept stirred at 25° C. for 2 h. Themixture was purified by a silica gel column to give desired product 17e(43 mg, 98%). HR-EIMS: m/z [M+H]⁺ 449.1241 (calcd. 449.1236). ¹H NMR(400 MHz, CDCl₃) δ 7.20 (1H, s, H-2), 7.10 (1H, s, H-5), 6.96 (1H, d,J=7.9 Hz, H-5′), 6.85 (1H, d, J=1.5 Hz, H-2′), 6.82 (1H, dd, J=7.9, 1.7Hz, H-6′), 6.09 (1H, br s, —OCH₂O—), 6.04 (1H, br s, —OCH₂O—), 5.24 (2H,s, H-9), 4.05 (3H, s, 3-OCH₃), 3.80 (3H, s, 4-OCH₃), 2.12-2.04 (1H, m,cyclopropanecarbonyl), 1.34-1.27 (2H, m, cyclopropanecarbonyl), 1.20(2H, dt, J=3.3, 8.0 Hz, cyclopropanecarbonyl). ¹³C NMR (100 MHz, CDCl₃)δ 172.2 (cyclopropanecarbonyl), 169.4 (C-9′), 152.3 (C-3), 150.4 (C-4),147.7 (C-3′), 147.6 (C-4′), 138.2 (C-7), 137.5 (C-7′), 130.7 (C-6),130.3 (C-1), 128.0 (C-1′), 126.3 (C-8′), 123.6 (C-6′), 119.1 (C-8),110.7 (C-2′), 108.3 (C-5′), 106.4 (C-5), 101.3 (C-2), 99.2 (—OCH₂O—),66.3 (C-9), 56.0 (3-OCH₃), 55.9 (4-OCH₃), 12.7 (cyclopropanecarbonyl),9.9 (cyclopropanecarbonyl).

Compound 17f

5 (38 mg, 0.1 mmol), Et₃N (30 mg, 0.3 mmol) and DMAP (catalyzed amount)were dissolved in 2 mL of dry CH₂Cl₂. Cyclopentanecarbonyl chloride (40mg, 0.3 mmol) was added to the mixture at 0° C. The reaction mixture wasallowed to warm up to 25° C. and kept stirred at 25° C. for 2 h. Themixture was purified by a silica gel column to give desired product 17f(45 mg, 97%). HR-EIMS: m/z [M+H]⁺ 477.1547 (calcd. 477.1549). ¹H NMR(400 MHz, CDCl₃) δ 7.12 (1H, s, H-2), 7.04 (1H, s, H-5), 6.90 (1H, d,J=7.9 Hz, H-5′), 6.79 (1H, d, J=1.5 Hz, H-2′), 6.76 (1H, dd, J=7.9, 1.7Hz, H-6′), 6.09 (1H, br s, —OCH₂O—), 6.04 (1H, br s, —OCH₂O—), 5.16 (2H,s, H-9), 3.96 (3H, s, 3-OCH₃), 3.74 (3H, s, 4-OCH₃), 3.16 (1H, p, J=8.0Hz, cyclopentanecarbonyl), 2.19-2.08 (2H, m, cyclopentanecarbonyl),2.07-1.96 (2H, m, cyclopentanecarbonyl), 1.80 (2H, dt, J=6.1, 17.0 Hz,cyclopentanecarbonyl), 1.70 (2H, tdd, J=2.2, 4.0, 11.4 Hz,cyclopentanecarbonyl). ¹³C NMR (100 MHz, CDCl₃) δ 173.8(cyclopentanecarbonyl), 169.4 (C-9′), 152.2 (C-3), 150.4 (C-4), 147.7(C-3′), 147.6 (C-4′), 138.2 (C-7), 137.5 (C-7′), 130.7 (C-6), 130.2(C-1), 128.0 (C-1′), 126.3 (C-8′), 123.7 (C-6′), 119.1 (C-8), 110.7(C-2′), 108.3 (C-5′), 106.4 (C-5), 101.3 (C-2), 99.2 (—OCH₂O—), 66.3(C-9), 56.0 (3-OCH₃), 55.9 (4-OCH₃), 43.8 (cyclopentanecarbonyl), 30.5(cyclopentanecarbonyl), 25.9 (cyclopentanecarbonyl).

Compound 17g

5 (38 mg, 0.1 mmol), Et₃N (30 mg, 0.3 mmol) and DMAP (catalyzed amount)were dissolved in 2 mL of dry CH₂Cl₂. 4-Fluorobenzoyl chloride (40 mg,0.3 mmol) was added to the mixture at 0° C. The reaction mixture wasallowed to warm up to 25° C. and kept stirred at 25° C. for 2 h. Themixture was purified by a silica gel column to give desired product 17g(45 mg, 90%). HR-EIMS: m/z [M+H]⁺ 503.1133 (calcd. 503.1142). ¹H NMR(400 MHz, CDCl₃) δ 8.37 (2H, dd, J=8.9, 5.3 Hz, 4-fluorobenzoyl), 7.29(2H, t, J=8.6 Hz, 4-fluorobenzoyl), 7.21 (1H, s, H-2), 7.15 (1H, s,H-5), 6.98 (1H, d, J=7.9 Hz, H-5′), 6.88 (1H, d, J=1.3 Hz, H-2′), 6.85(1H, dd, J=7.9, 1.7 Hz, H-6′), 6.09 (1H, br s, —OCH₂O—), 6.04 (1H, br s,—OCH₂O—), 5.29 (2H, s, H-9), 3.95 (3H, s, 3-OCH₃), 3.82 (3H, s, 4-OCH₃).¹³C NMR (100 MHz, CDCl₃) δ 169.7 (C-9′), 168.4 (4-fluorobenzoyl), 165.8(4-fluorobenzoyl), 163.1 (4-fluorobenzoyl), 152.8 (C-3), 150.9 (C-4),148.1 (C-3′), 148.0 (C-4′), 138.5 (C-7), 138.3 (C-7′), 133.6(4-fluorobenzoyl), 133.5 (4-fluorobenzoyl), 131.2 (C-1), 130.8 (C-6),128.3 (C-1′), 126.7 (C-8′), 124.1 (C-6′), 119.6 (C-8), 116.9(4-fluorobenzoyl), 116.7 (4-fluorobenzoyl), 111.1 (C-2′), 108.7 (C-5′),106.9 (C-5), 101.7 (C-2), 99.5 (—OCH₂O—), 66.8 (C-9), 56.4 (3-OCH₃),56.3 (4-OCH₃).

Compound 17h

5 (38 mg, 0.1 mmol), Et₃N (30 mg, 0.3 mmol) and DMAP (catalyzed amount)were dissolved in 2 mL of dry CH₂Cl₂. 4-Methoxybenzoyl chloride (51 mg,0.3 mmol) was added to the mixture at 0° C. The reaction mixture wasallowed to warm up to 25° C. and kept stirred at 25° C. for 2 h. Themixture was purified by a silica gel column to give desired product 17h(45 mg, 90%). HR-EIMS: m/z [M+H]⁺ 515.1338 (calcd. 515.1342). ¹H NMR(400 MHz, CDCl₃) δ 8.33-8.25 (2H, m, 4-methoxybenzoyl), 7.26 (1H, s,H-2), 7.14 (1H, s, H-5), 7.10-7.03 (2H, m, 4-methoxybenzoyl), 6.97 (1H,d, J=7.9 Hz, H-5′), 6.88 (1H, d, J=1.4 Hz, H-2′), 6.86 (1H, dd, J=7.9,1.7 Hz, H-6′), 6.09 (1H, d, J=1.4 Hz, —OCH₂O—), 6.04 (1H, d, J=1.4 Hz,—OCH₂O—), 5.29 (2H, s, H-9), 3.94 (3H, s, 3-OCH₃), 3.93 (3H, s,4-methoxybenzoyl), 3.81 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, CDCl₃) δ169.4 (C-9′), 164.7 (4-methoxybenzoyl), 163.4 (4-methoxybenzoyl), 152.3(C-3), 150.4 (C-4), 147.7 (C-3′), 147.6 (C-4′), 138.5 (C-7), 137.5(C-7′), 132.7 (4-methoxybenzoyl), 130.7 (C-6), 130.6 (C-1), 128.0(C-1′), 126.6 (C-8′), 123.7 (C-6′), 120.3 (4-methoxybenzoyl), 119.2(C-8), 114.4 (4-methoxybenzoyl), 110.7 (C-2′), 108.3 (C-5′), 106.4(C-5), 101.3 (C-2), 99.4 (—OCH₂O—), 66.6 (C-9), 56.0 (3-OCH₃), 55.9(4-OCH₃), 55.7 (4-methoxybenzoyl).

Compound 18

Under nitrogen, diphyllin (5, 760 mg, 2 mmol) and DMAP (488 mg, 4 mmol)were dissolved in 10 mL dry CH₂Cl₂. The solution was cooled to 0° C.Then trifluoromethanesulfonic anhydride (Tf₂O) (0.5 mL, 2.4 mmol) wasadded slowly via syringe over 5 min. The reaction was allowed to warm upto r.t. and kept stirred at r.t. for 4 h. TLC showed that the reactionhad completed, and a new spot formed. The mixture was concentrated invacuo to give a yellow solid, which was washed with EtOH to give desiredproduct 18 (0.97 g, 95%). HR-EIMS: m/z [M+H]⁺ 513.0455 (calcd.513.0467). ¹H NMR (400 MHz, DMSO-d₆) δ 7.35 (1H, s, H-2), 7.11 (1H, s,H-5), 7.08 (1H, d, J=7.9 Hz, H-5′), 7.01 (1H, d, J=1.6 Hz, H-2′), 6.87(1H, dd, J=7.9, 1.7 Hz, H-6′), 6.15 (2H, s, —OCH₂O—), 5.56 (2H, s, H-9),3.99 (3H, s, 3-OCH₃), 3.72 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ167.7 (C-9′), 153.1 (C-3), 150.6 (C-4), 147.5 (C-3′), 147.1 (C-4′),139.7 (C-7), 135.2 (C-7′), 131.6 (C-6), 130.2 (C-1), 126.8 (C-1′), 125.2(C-8′), 123.6 (C-6′), 119.7 (OTf), 119.4 (C-8), 110.7 (C-2′), 108.1(C-5′), 106.2 (C-5), 101.3 (C-2), 98.7 (—OCH₂O—), 65.4 (C-9), 55.9(3-OCH₃), 55.5 (4-OCH₃).

Compound 19a

In an oven dried round bottom flask under an atmosphere of nitrogen wasplaced [Pd(dppf)Cl₂] CH₂Cl₂ complex (244 mg, 0.025 mmol), B₂Pin₂ (191mg, 0.75 mmol), KOAc (147 mg, 1.5 mmol) and 18 (256 mg, 0.5 mmol). Tothe flask was added freshly distilled 1,4-dioxane (2 mL). The mixturewas degassed with N₂. The flask was equipped with a reflux condenser andheated at 100° C. for 3 h under a nitrogen atmosphere. The mixture wasconcentrated in vacuo to give a yellow solid, which was washed with EtOHto give desired product 19a (229.5 mg, 85%). HR-EIMS: m/z [M+H]⁺491.3626 (calcd. 491.1882). ¹H NMR (400 MHz, DMSO-d₆) δ 7.51 (1H, s,H-2), 7.04 (1H, d, J=7.9 Hz, H-5′), 6.98 (1H, s, H-5), 6.92 (1H, d,J=1.6 Hz, H-2′), 6.79 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.13 (2H, br s,—OCH₂O—), 5.43 (2H, s, H-9), 3.94 (3H, s, 3-OCH₃), 3.66 (3H, s, 4-OCH₃),1.06 (12H, s, Me). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.5 (C-9′), 151.5(C-3), 149.8 (C-4), 147.0 (C-3′, C-4′), 139.9 (C-7), 138.4 (C-7′), 133.0(C-6), 128.4 (C-1′), 127.8 (C-1), 123.5 (C-6′), 118.9 (C-8), 117.9(C-8′), 110.7 (C-2′), 108.0 (C-5′), 106.7 (C-5), 105.0 (C-2), 101.2(—OCH₂O—), 79.1 (C—O), 73.6 (C—O), 68.1 (C-9), 55.9 (3-OCH₃), 55.2(4-OCH₃), 25.0 (CH₃).

Compound 19b

An oven-dried 10-mL flask was charged with Pd(dppf)Cl₂ (3.7 mg, 0.005mmol) and 18 (51 mg, 0.1 mmol). The flask was capped and then backfilledwith N2. Toluene (2.00 mL) was added via syringe, followed by theaddition of AlMe₃ (0.15 mL, 0.3 mmol). The reaction mixture was heatedto 100° C. for 10 h. After cooling to r.t., the reaction solution wasquenched with water and extracted with CH₂Cl₂. The obtained solution wasconcentrated and purified by prep-TLC to give desired product 19b (25mg, 66%). HR-EIMS: m/z [M+H]+ 379.1156 (calcd. 379.1182). ¹H NMR (400MHz, DMSO-d₆) δ 7.40 (1H, s, H-2), 7.04 (1H, d, J=7.9 Hz, H-5′), 7.01(1H, s, H-5), 6.88 (1H, d, J=1.6 Hz, H-2′), 6.77 (1H, dd, J=7.9, 1.7 Hz,H-6′), 6.12 (2H, s, —OCH₂O—), 5.44 (s, 2H, H-9), 3.99 (3H, s, 3-OCH₃),3.65 (3H, s, 4-OCH₃), 2.57 (3H, s, Me). ¹³C NMR (100 MHz, DMSO-d₆) δ169.9 (C-9′), 151.3 (C-3), 149.3 (C-4), 147.0 (C-3′, C-4′), 138.2 (C-7),136.5 (C-7′), 131.5 (C-6), 128.6 (C-1′), 128.1 (C-1), 125.6 (C-8′),123.6 (C-6′), 117.4 (C-8), 110.8 (C-2′), 108.0 (C-5′), 105.9 (C-5),102.8 (C-2), 101.2 (—OCH₂O—), 67.9 (C-9), 55.8 (3-OCH₃), 55.2 (4-OCH₃),14.3 (CH₃).

Compound 19c

An oven-dried 10-mL flask was charged with Pd(dppf)₂Cl₂ (7.31 mg, 0.01mmol), Na₂CO₃ (65 mg, 0.6 mmol), 18 (102 mg, 0.2 mmol) andpyridine-4-boronic acid (52 mg, 0.4 mmol). The flask was capped and thenbackfilled with N₂. Dioxane and H₂O (2/0.4 mL) was added via syringe.The reaction mixture was heated to 100° C. for 10 h. After cooling tor.t., the reaction solution was quenched with water and extracted withCH₂Cl₂. The obtained solution was concentrated and purified by prep-TLCto give desired product 19c (22 mg, 50%). HR-EIMS: m/z [M+H]⁺ 442.1300(calcd. 442.1291). ¹H NMR (400 MHz, DMSO-d₆) δ 8.79 (2H, d, J=5.8 Hz,pyridine), 7.62 (2H, s, H-2, pyridine), 7.09 (1H, s, pyridine), 7.08(1H, d, J=8.1 Hz, H-5′), 7.01 (1H, s, H-5), 6.97 (1H, d, J=1.5 Hz,H-2′), 6.85 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.14 (2H, s, —OCH₂O—), 5.27(2H, s, H-9), 3.72 (3H, s, 3-OCH₃), 3.68 (3H, s, 4-OCH₃). ¹³C NMR (100MHz, DMSO-d₆) δ 169.2 (C-9′), 151.8 (C-3), 150.4 (pyridine), 149.7(C-4), 147.1 (C-3′), 147.0 (C-4′), 143.8 (pyridine), 138.9 (C-7), 138.2(C-7′), 129.9 (C-6), 128.5 (C-1), 128.0 (C-1′), 124.5 (pyridine, C-8′),123.5 (C-6′), 117.9 (C-8), 110.6 (C-2′), 108.1 (C-5′), 105.8 (C-5),103.3 (C-2), 101.2 (—OCH₂O—), 67.5 (C-9), 55.4 (3-OCH₃), 55.3 (4-OCH₃).

Compound 19d

An oven-dried 10-mL flask was charged with Pd(OAc)₂ (1.1 mg, 0.01 mmol),PPh₃ (2.6 mg, 0.02 mmol) and 18 (0.1 mmol). The flask was capped andthen backfilled with argon (this was repeated two additional times). DMF(1.00 mL) was added via syringe, followed by the addition of Et₃N (43mL, 0.3 mmol) and HCO₂H (8 μL, 0.2 mmol) in a like manner. The reactionmixture was heated to 100° C. for 10 h. After cooling to r.t., thereaction solution was quenched with water and extracted with CH₂Cl₂. Theobtained solution was concentrated and purified by prep-TLC to givedesired product 19d (35 mg, 98%). HR-EIMS: m/z [M+H]⁺ 365.1010 (calcd.365.1025). ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 (1H, s, H-7), 7.51 (1H, s,H-2), 7.05 (1H, d, J=7.9 Hz, H-5′), 6.99 (1H, s, H-5), 6.92 (1H, d,J=1.6 Hz, H-2′), 6.80 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.13 (2H, s,—OCH₂O—), 5.43 (2H, s, H-9), 3.94 (3H, s, 3-OCH₃), 3.66 (3H, s, 4-OCH₃).¹³C NMR (100 MHz, DMSO-d₆) δ 169.5 (C-9′), 151.5 (C-3), 149.8 (C-4),147.0 (C-3′, C-4′), 139.9 (C-7), 138.3 (C-7′), 132.9 (C-6), 128.4(C-1′), 127.8 (C-1), 123.5 (C-6′), 118.8 (C-8), 117.9 (C-8′), 110.7(C-2′), 108.0 (C-5′), 106.7 (C-5), 105.0 (C-2), 101.2 (—OCH₂O—), 68.1(C-9), 56.0 (3-OCH₃), 55.8 (4-OCH₃).

Compound 19e

An oven-dried 10-mL flask was charged with Pd(OAc)₂ (1.12 mg, 0.005mmol), BINAP (5 mg, 0.008 mmol), Cs₂CO₃ (49 mg, 0.15 mmol) and 18 (51mg, 0.1 mmol). The flask was capped and then backfilled with N₂. Asolution of 4-aminobenzotrifluoride (32 mg, 0.2 mmol) in PhMe was addedvia syringe. The reaction mixture was heated to 100° C. for 10 h. Aftercooling to r.t., the reaction solution was quenched with water andextracted with CH₂Cl₂. The obtained solution was concentrated andpurified by prep-TLC to give desired product 19e (45 mg, 86%). HR-EIMS:m/z [M+H]⁺ (calcd. 524.1321). ¹H NMR (400 MHz, CDCl₃) δ 7.44 (2H, d,J=8.5 Hz, benzotrifluoride), 7.32 (1H, s, H-2), 7.15 (1H, s, H-5), 6.93(1H, d, J=7.9 Hz, H-5′), 6.86 (1H, d, J=1.5 Hz, H-2′), 6.83 (1H, dd,J=7.9, 1.7 Hz, H-6′), 6.70 (2H, d, J=8.4 Hz, benzotrifluoride), 6.06(1H, br s, —OCH₂O—), 5.96 (1H, br s, —OCH₂O—), 5.13 (2H, s, H-9), 3.88(3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, CDCl₃) δ 170.6(C-9′), 152.6 (C-3), 150.7 (C-4), 148.2 (benzotrifluoride), 148.1(C-3′), 148.0 (C-4′), 138.3 (C-7), 136.0 (C-7′), 130.9(benzotrifluoride), 130.8 (C-6), 128.6 (C-8′), 128.5 (C-1), 127.3(C-1′), 124.0 (C-6′), 119.4 (C-8), 114.3 (benzotrifluoride), 111.0(C-2′), 108.6 (C-5′), 107.1 (C-5), 101.9 (C-2), 101.8 (—OCH₂O—), 67.8(C-9), 56.5 (3-OCH₃), 56.3 (4-OCH₃).

Compound 19f

An oven-dried 10-mL flask was charged with Pd(OAc)₂ (1.12 mg, 0.005mmol), BINAP (5 mg, 0.008 mmol), Cs₂CO₃ (49 mg, 0.15 mmol) and 18 (51mg, 0.1 mmol). The flask was capped and then backfilled with N₂. Asolution of aminobenzene (19 mg, 0.2 mmol) in PhMe was added viasyringe. The reaction mixture was heated to 100° C. for 10 h. Aftercooling to r.t., the reaction solution was quenched with water andextracted with CH₂Cl₂. The obtained solution was concentrated andpurified by prep-TLC to give desired product 19g (38.7 mg, 85%).HR-EIMS: m/z [M+H]+(calcd. 456.1447). ¹H NMR (400 MHz, CDCl₃) δ 7.40(1H, s, H-2), 7.22 (2H, dd, J=8.5, 7.4 Hz, aminobenzene), 7.14 (1H, s,H-5), 6.93 (1H, d, J=7.9 Hz, H-5′), 6.89-6.87 (1H, m, aminobenzene),6.86 (1H, d, J=1.5 Hz, H-2′), 6.83 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.72(2H, dd, J=1.0, 8.5 Hz, aminobenzene), 6.06 (1H, br s, —OCH₂O—), 5.98(1H, br s, —OCH₂O—), 5.07 (2H, s, H-9), 3.89 (3H, s, 3-OCH₃), 3.80 (3H,s, 4-OCH₃). ¹³C NMR (100 MHz, CDCl₃) δ 170.4 (C-9′), 151.9 (C-3), 150.1(C-4), 147.6 (C-3′), 147.6 (C-4′), 144.6 (C-7), 136.4 (C-7′), 133.9(aminobenzene), 130.4 (C-6), 129.8 (C-1), 129.7 (C-8′), 129.5(aminobenzene), 128.6 (C-1′), 123.7 (C-6′), 120.2 (C-8), 119.2(aminobenzene), 115.6 (aminobenzene), 110.8 (C-2′), 108.2 (C-5′), 106.7(C-5), 101.7 (C-2), 101.3 (—OCH₂O—), 67.7 (C-9), 56.1 (3-OCH₃), 55.9(4-OCH₃).

Compound 19g

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (6.22 mg, 0.005mmol) and 18 (51 mg, 0.1 mmol). The flask was capped and then backfilledwith N₂. THF (2.00 mL) was added via syringe, followed by the additionof Et₂Zn. The reaction mixture was heated to 100° C. for 10 h. Aftercooling to r.t., the reaction solution was quenched with water andextracted with CH₂Cl₂. The obtained solution was concentrated andpurified by prep-TLC to give desired product 19g (25 mg, 64%). HR-EIMS:m/z [M+H]⁺ 393.1324 (calcd. 393.1338). ¹H NMR (400 MHz, DMSO-d₆) δ 7.43(1H, s, H-2), 7.04 (1H, d, J=8.1 Hz, H-5′), 7.02 (1H, s, H-5), 6.90 (1H,d, J=1.3 Hz, H-2′), 6.77 (1H, dd, J=7.7, 1.3 Hz, H-6′), 6.12 (2H, s,—OCH₂O—), 5.47 (2H, s, H-9), 3.99 (3H, s, 3-OCH₃), 3.65 (3H, s, 4-OCH₃),3.03 (2H, q, J=7.3 Hz, Et), 1.28 (3H, t, J=7.5 Hz, Et). ¹³C NMR (100MHz, DMSO-d₆) δ 169.9 (C-9′), 151.5 (C-3), 149.3 (C-4), 147.0 (C-3′),146.9 (C-4′), 137.7 (C-7), 136.7 (C-7′), 131.7 (C-6), 130.6 (C-1), 128.7(C-8′), 128.5 (C-1′), 123.6 (C-6′), 117.6 (C-8), 110.8 (C-2′), 108.1(C-5′), 106.1 (C-5), 102.5 (C-2), 101.2 (—OCH₂O—), 67.6 (C-9), 55.8(3-OCH₃), 55.2 (4-OCH₃), 21.5 (Et), 13.9 (Et).

Compound 19h

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (6.22 mg, 0.005mmol), K₂CO₃ (40 mg, 0.3 mmol), 18 (51 mg, 0.1 mmol) andthiophene-2-boronic acid (26 mg, 0.2 mmol). The flask was capped andthen backfilled with N₂. DMF was added via syringe. The reaction mixturewas heated to 100° C. for 10 h. After cooling to r.t., the reactionsolution was quenched with water and extracted with CH₂Cl₂. The obtainedsolution was concentrated and purified by prep-TLC to give desiredproduct 19h (38 mg, 92%). HR-EIMS: m/z [M+H]⁺ 447.0889 (calcd.447.0902). ¹H NMR (400 MHz, DMSO-d₆) δ 7.87 (1H, dd, J=5.1, 1.2 Hz,thiophene), 7.42 (1H, dd, J=3.5, 1.2 Hz, thiophene), 7.41 (1H, s, H-2),7.33 (1H, dd, J=5.1, 3.5 Hz, thiophene), 7.07 (1H, d, J=7.9 Hz, H-5′),7.06 (1H, s, H-5), 6.95 (1H, d, J=1.5 Hz, H-2′), 6.83 (1H, dd, J=7.9,1.7 Hz, H-6′), 6.13 (2H, br s, —OCH₂O—), 5.32 (2H, s, H-9), 3.77 (3H, s,3-OCH₃), 3.67 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.2(C-9′), 151.7 (C-3), 149.6 (C-4), 147.0 (C-3′, C-4′), 139.6 (C-7), 138.7(thiophene), 135.4 (C-7′), 131.4 (C-6), 129.0 (C-1), 128.6 (C-1′), 128.1(thiophene), 128.1 (thiophene), 128.1 (thiophene), 123.6 (C-6′), 123.5(C-8′), 118.0 (C-8), 110.6 (C-2′), 108.1 (C-5′), 105.7 (C-5), 103.7(C-2), 101.2 (—OCH₂O—), 67.9 (C-9), 55.4 (3-OCH₃), 55.3 (4-OCH₃).

Compound 19i

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (6.22 mg, 0.005mmol), K₂CO₃ (42 mg, 0.3 mmol), 18 (51 mg, 0.1 mmol) andpyridine-3-boronic acid (25 mg, 0.2 mmol). The flask was capped and thenbackfilled with N₂. 2 mL DMF was added via syringe. The reaction mixturewas heated to 100° C. for 10 h. After cooling to r.t., the reactionsolution was quenched with water and extracted with CH₂Cl₂. The obtainedsolution was concentrated and purified by prep-TLC to give desiredproduct 19i (30 mg, 75%). HR-EIMS: m/z [M+H]⁺ 442.1293 (calcd.442.1291). ¹H NMR (400 MHz, DMSO-d₆) δ 8.80 (2H, s, pyridine), 8.09 (1H,s, pyridine), 7.71 (1H, s, H-2), 7.09 (1H, s, pyridine), 7.08 (2H, d,J=8.3 Hz, H-5′), 6.97 (1H, d, J=4.2 Hz, H-2′), 6.95 (1H, s, H-5), 6.85(1H, dd, J=8.6, 4.2 Hz, H-6′), 6.14 (2H, br s, —OCH₂O—), 5.36-5.19 (2H,m, H-9), 3.71 (3H, s, 3-OCH₃), 3.68 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz,DMSO-d₆) δ 169.3 (C-9′), 151.7 (C-3), 149.6 (C-4, pyridine), 147.1(C-3′, C-4′), 139.0 (C-7, pyridine), 138.7 (pyridine), 138.0 (C-7′),130.8 (C-6, pyridine), 128.5 (C-1′, C-1), 128.1 (pyridine), 127.1(C-8′), 123.5 (C-6′), 117.8 (C-8), 110.6 (C-2′), 108.1 (C-5′), 105.7(C-5), 103.4 (C-2), 101.2 (—OCH₂O—), 67.5 (C-9), 55.4 (3-OCH₃), 55.3(4-OCH₃).

Compound 19j

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (6.22 mg, 0.005mmol), K₂CO₃ (42 mg, 0.3 mmol), 18 (51 mg, 0.1 mmol) andthiophene-3-boronic acid (26 mg, 0.2 mmol). The flask was capped andthen backfilled with N₂. DMF 2 mL was added via syringe. The reactionmixture was heated to 100° C. for 10 h. After cooling to r.t., thereaction solution was quenched with water and extracted with CH₂Cl₂. Theobtained solution was concentrated and purified by prep-TLC to givedesired product 19j (22 mg, 50%). HR-EIMS: m/z [M+H]⁺ 447.0867 (calcd.447.0902). ¹H NMR (400 MHz, DMSO-d₆) δ 7.91 (1H, d, J=1.4 Hz,thiophene), 7.84 (1H, d, J=4.7 Hz, thiophene), 7.44 (1H, d, J=4.9 Hz,thiophene), 7.27 (1H, s, H-2), 7.06 (1H, d, J=7.9 Hz, H-5′), 7.05 (1H,s, H-5), 6.95 (1H, d, J=1.0 Hz, H-2′), 6.83 (1H, dd, J=7.8, 1.3 Hz,H-6′), 6.13 (2H, s, —OCH₂O—), 5.32 (2H, s, H-9), 3.76 (3H, s, 3-OCH₃),3.66 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO) δ 170.0 (C-9′), 151.9(C-3), 150.0 (C-4), 147.5 (C-3′, C-4′), 139.0 (C-7), 138.3 (C-7′), 136.0(thiophene), 131.3 (C-6), 129.2 (C-1), 129.0 (thiophene), 128.8 (C-1′),127.8 (C-8′), 126.6 (thiophene), 125.9 (thiophene), 123.9 (C-6′), 118.4(C-8), 111.1 (C-2′), 108.5 (C-5′), 106.1 (C-5), 104.4 (C-2), 101.7(—OCH₂O—), 68.4 (C-9), 55.8 (3-OCH₃), 55.7 (4-OCH₃).

Compound 19k

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (6.22 mg, 0.005mmol), Na₂CO₃ (32 mg, 0.3 mmol), 18 (51 mg, 0.1 mmol) andfuran-2-boronic acid (26 mg, 0.2 mmol). The flask was capped and thenbackfilled with N₂. Dioxane and H₂O (2/0.4 mL) was added via syringe.The reaction mixture was heated to 100° C. for 10 h. After cooling tor.t., the reaction solution was quenched with water and extracted withCH₂Cl₂. The obtained solution was concentrated and purified by prep-TLCto give desired product 19k (30 mg, 70%). HR-EIMS: m/z [M+H]⁺ 431.1098(calcd. 431.1131). ¹H NMR (400 MHz, DMSO-d₆) δ 7.99 (1H, dd, J=1.8, 0.7Hz, furan), 7.73 (1H, s, H-2), 7.14 (1H, dd, J=3.4, 0.6 Hz, furan), 7.07(1H, d, J=7.9 Hz, H-5′), 7.06 (1H, s, H-5), 6.95 (d, 1H, J=1.5 Hz,furan), 6.82 (1H, dd, J=7.9, 1.8 Hz, H-6′), 6.82 (1H, d, J=1.9 Hz,H-2′), 6.14 (2H, s, —OCH₂O—), 5.51 (2H, s, H-9), 3.92 (3H, s, 3-OCH₃),3.68 (3H, s, 4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.2 (C-9′), 152.0(C-3), 149.6 (C-4), 148.9 (furan), 147.1 (C-3′), 147.0 (C-4′), 143.9(furan), 138.5 (C-7), 138.4 (C-7′), 129.1 (C-6), 128.9 (C-1), 128.2(C-1′), 123.5 (C-6′), 119.8 (C-8), 118.3 (C-8′), 112.1 (furan), 111.8(furan), 110.7 (C-2′), 108.1 (C-5′), 105.9 (C-5), 104.1 (C-2), 101.3(—OCH₂O—), 68.6 (C-9), 55.5 (3-OCH₃), 55.2 (4-OCH₃).

Compound 191

An oven-dried 10-mL flask was charged with Pd(PPh₃)₄ (24 mg, 0.02 mmol),Na₂CO₃ (120 mg, 1.2 mmol), 18 (204 mg, 0.4 mmol),3,6-dihydro-2H-pyridine-1-N-Boc-4-boronic acid, and pinacol ester (186mg, 0.6 mmol). The flask was capped and then backfilled with N2. Dioxaneand water (4/0.8 mL) were added via syringe. The reaction mixture washeated to 80° C. for 10 h. After cooling to r.t., the reaction solutionwas quenched with water and extracted with CH₂Cl₂. The mixture waspurified by a silica gel column to give desired product 19l (69 mg,27%). HR-EIMS: m/z [M+H]⁺ 546.2109 (calcd. 546.2128).1H NMR (400 MHz,CDCl₃) δ 7.12 (1H, s, H-2), 7.11 (1H, s, H-5), 6.96 (1H, d, J=7.9 Hz,H-5′), 6.84 (1H, br s, H-2′), 6.81 (1H, d, J=8.1 Hz, H-6′), 6.09 (1H, brs, —OCH₂O—), 6.04 (1H, br s, —OCH₂O—), 5.88 (1H, s,3,6-dihydro-2H-pyridine), 5.29 (2H, s, H-9), 4.42-4.05 (2H, m,3,6-dihydro-2H-pyridine) 3.99 (3H, s, 3-OCH₃), 3.80 (3H, s, 4-OCH₃),3.69 (2H, s, 3,6-dihydro-2H-pyridine), 2.59-2.29 (2H, m,3,6-dihydro-2H-pyridine), 1.53 (9H, s, Boc). ¹³C NMR (100 MHz, CDCl₃) δ170.1 (C-9′), 155.1 (Boc), 151.7 (C-3), 149.8 (C-4), 147.6 (C-3′), 147.6(C-4′), 138.9 (C-7), 136.9 (C-7′), 131.9 (3,6-dihydro-2H-pyridine),130.7 (C-6), 129.4 (C-1), 128.4 (C-1′), 124.5 (3,6-dihydro-2H-pyridine),123.6 (C-6′), 123.5 (C-8′), 118.3 (C-8), 108.3 (C-2′, C-5′), 106.5(C-5), 103.3 (C-2), 101.3 (—OCH₂O—), 80.2 (Boc), 67.7(3,6-dihydro-2H-pyridine), 67.1 (C-9), 55.9 (3-OCH₃), 55.9 (4-OCH₃),53.5 (3,6-dihydro-2H-pyridine), 29.4 (3,6-dihydro-2H-pyridine), 28.5(Boc).

Compound 19m

An oven-dried 10-mL flask was charged with Pd₂(dba)₃ (4.58 mg, 0.005mmol), XantPhos (2.89 mg, 0.005 mmol), Cs₂CO₃ (65 mg, 0.2 mmol) and 18(51 mg, 0.1 mmol). The flask was capped and then backfilled with N₂. Asolution of morpholine (17 mg, 0.2 mmol) in toluene was added viasyringe. The reaction mixture was heated to 100° C. for 10 h. Aftercooling to r.t., the reaction solution was quenched with water andextracted with CH₂Cl₂. The obtained solution was concentrated andpurified by prep-TLC to give desired product 19m (20 mg, 40%). HR-EIMS:m/z [M+H]⁺ (calcd. 450.1553). ¹H NMR (400 MHz, CDCl₃) δ 7.79 (1H, s,H-2), 7.07 (1H, s, H-5), 6.96 (1H, d, J=7.9 Hz, H-5′), 6.82 (1H, d,J=1.4 Hz, H-2′), 6.79 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.09 (1H, br s,—OCH₂O—), 6.05 (1H, br s, —OCH₂O—), 5.52 (2H, s, H-9), 4.07 (3H, s,3-OCH₃), 4.05-4.01 (2H, m, morpholine), 3.93 (2H, td, J=10.9, 2.2 Hz,morpholine), 3.80 (3H, s, 4-OCH₃), 3.28 (2H, td, J=11.3, 2.9 Hz,morpholine), 3.13 (2H, d, J=11.8 Hz, morpholine). ¹³C NMR (100 MHz,CDCl₃) δ 169.6 (C-9′), 151.8 (C-3), 150.1 (C-4), 147.5 (C-3′, C-4′),142.2 (C-7), 139.4 (C-7′), 136.8 (C-6), 132.1 (C-1), 130.5 (C-8′), 129.9(C-1′), 128.6 (C-8), 123.5 (C-6′), 110.6 (C-2′), 108.2 (C-5′), 106.5(C-5), 102.6 (C-2), 101.2 (—OCH₂O—), 67.8 (morpholine), 67.3 (C-9), 55.9(3-OCH₃), 55.9 (4-OCH₃), 51.5 (morpholine).

Compound 19n

A solution of 18 (256 mg, 0.5 mmol), tributyl (1-ethoxyvinyl) stannane(216 mg, 0.6 mmol) and 5 mL toluene was degassed for 30 min whilestirring. PdCl₂(PPh₃)₂ (34 mg, 0.05 mmol) was then added, the solutionwas heated to 100° C. overnight with stirring. 10 mL of 50% HCl (aq.)was added to the reaction mixture and the solution was stirred for 6 h,followed by filtration through celite that effectively removed thepalladium black. The mixture was purified by a silica gel column to givedesired product 19n (180 mg, 88%). HR-EIMS: m/z [M+H]⁺ 393.1324 (calcd.393.1338). ¹H NMR (400 MHz, DMSO-d₆) δ 7.64 (1H, s, H-2), 7.07 (1H, d,J=7.9 Hz, H-5′), 7.06 (1H, s, H-5), 6.96 (1H, d, J=1.6 Hz, H-2′), 6.82(1H, dd, J=7.9, 1.7 Hz, H-6′), 6.14 (2H, s, —OCH₂O—), 5.55 (2H, s, H-9),3.97 (31H, s, 3-OCH₃), 3.68 (3H, s, 4-OCH₃), 2.76 (3H, s, CH₃). ¹³C NMR(100 MHz, DMSO-d₆) δ 202.0 (Me-C═O), 168.9 (C-9′), 152.4 (C-3), 149.7(C-4), 147.3 (C-3′), 147.1 (C-4′), 141.6 (C-7), 139.6 (C-7′), 129.4(C-6), 129.1 (C-1), 129.0 (C-8′), 127.9 (C-1′), 123.5 (C-6′), 118.0(C-8), 110.6 (C-2′), 108.2 (C-5′), 106.0 (C-5), 104.4 (C-2), 101.4(—OCH₂O—), 68.6 (C-9), 55.9 (3-OCH₃), 55.3 (4-OCH₃), 31.8 (CH₃).

Compound 19o

An oven-dried 25-mL flask was charged with Pd(PPh₃)₂Cl₂ (17.5 mg, 0.025mmol), CuI (14.3 mg, 0.075 mmol) and 18 (255 mg, 0.5 mmol). The flaskwas capped and then backfilled with N₂. A solution oftrimethylsilylacetylene (100 mg, 1 mmol) and Et₃N (150 mg, 1.5 mmol) in5 mL DMF was added via syringe. The reaction mixture was heated to 80°C. for 10 h. After cooling to r.t., the reaction solution was quenchedwith water and extracted with CH₂Cl₂. The obtained solution wasconcentrated and purified by a silica gel column to give desired product19o (207 mg, 90%). HR-EIMS: m/z [M+H]⁺ 461.1404 (calcd. 461.1420). ¹HNMR (400 MHz, CDCl₃) 37.62 (1H, s, H-2), 7.06 (1H, s, H-5), 6.90 (1H, d,J=7.9 Hz, H-5′), 6.78 (1H, d, J=1.5 Hz, H-2′), 6.75 (1H, dd, J=7.9, 1.7Hz, H-6′), 6.03 (1H, br s, —OCH₂O—), 5.99 (1H, br s, —OCH₂O—), 5.34 (2H,s, H-9), 4.02 (3H, s, 3-OCH₃), 3.75 (3H, s, 4-OCH₃), 0.29 (9H, s, TMS).¹³C NMR (100 MHz, CDCl₃) δ 169.9 (C-9′), 152.6 (C-3), 150.3 (C-4), 147.9(C-3′), 147.7 (C-4′), 143.2 (C-7), 140.1 (C-7′), 133.3 (C-6), 128.9(C-1′), 128.1 (C-1), 123.7 (C-8′, C-6′), 118.5 (C-8), 110.7 (C-2′),108.4 (C-5′), 106.5 (C-5), 106.2 (C≡C), 104.5 (C≡C), 101.5 (C-2), 98.8(—OCH₂O—), 68.3 (C-9), 56.1 (3-OCH₃), 56.0 (4-OCH₃), 0.23 (TMS).

Compound 19p

A 25 mL round-bottomed flask (equipped with thermocouple, overheadstirring and a nitrogen inlet) was charged with cesium carbonate (74.1mg, 0.228 mmol), palladium(II) acetate (0.7 mg, 0.003 mmol), and4,5-bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene (Xantphos, 2.82 mg,0.005 mmol). The flask was evacuated and backfilled with nitrogen 3×.

Dioxane (2 mL) was added followed by another set of three cycles ofevacuation and backfilling with nitrogen. The canary yellow slurry wasstirred at r.t. for 10 min, and triethylamine (0.68 μL, 0.005 mmol) wasadded. Continued stirring for 10 min at r.t. gradually changed the colorfrom yellow to red. 18 (83 mg, 0.162 mmol) and benzophenone imine (32.7μL, 0.195 mmol) were added in 1 mL of dioxane via syringe (1 mL wash).The reaction mixture was warmed to an internal temperature of 100° C.for 12 h. Cooling the reaction mixture to ambient temperature wasfollowed by pouring the slurry into 10 mL of ethyl acetate. The organicswere washed with 2×50 mL of water, and the ethyl acetate solution wasconcentrated in vacuo. The mixture was purified by flash chromatographyon silica gel, eluting with 1:1 n-hexane:ethyl acetate, to give 19p (50mg, 57%) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.88 (2H, dt,J=7.1, 1.4 Hz, phenyl), 7.64-7.55 (1H, m, H-2), 7.50 (2H, ddd, J=8.1,6.5, 1.4 Hz, phenyl), 7.37-7.30 (2H, m, phenyl), 7.25-7.18 (2H, m,phenyl), 7.16-7.10 (2H, m, phenyl), 7.10 (1H, s, H-5), 6.94 (1H, dd,J=7.9, 0.5 Hz, H-5′), 6.82 (1H, dd, J=1.7, 0.5 Hz, H-2′), 6.79 (1H, dd,J=7.9, 1.7 Hz, H-6′), 6.08 (1H, d, J=1.5 Hz, —OCH₂O—), 6.03 (1H, d,J=1.5 Hz, —OCH₂O—), 5.29 (2H, s, H-9), 3.98 (3H, s, 3-OCH₃), 3.82 (3H,s, 4-OCH₃). ¹³C NMR (100 MHz, CDCl₃) δ 170.1 (C-9′), 151.3 (C-3), 149.8(C-4), 147.2 (C-3′), 147.2 (C-4′), 135.7 (C-7), 134.6 (C-7′), 131.5(C-6), 130.2 (C-1), 129.9 (C-1′), 129.8 (phenyl), 129.3 (phenyl), 128.4(phenyl), 128.3 (phenyl), 128.2 (phenyl), 128.1 (phenyl), 126.0 (C-6′),123.6 (C-8′), 118.3 (C-8), 110.8 (C-2′), 107.9 (C-5′), 106.3 (C-5),101.7 (C-2), 101.0 (—OCH₂O—), 67.0 (C-9), 56.0 (3-OCH₃), 55.6 (4-OCH₃).

Compound 20

To a solution of 19n (41 mg, 0.1 mmol) in 1 mL MeOH was added NaBH₄ (6mg, 0.15 mmol). The mixture was stirred at r.t. for 1 h. TLC showed thatthe reaction had finished with one spot and the mixture was purified byprep-TLC to give desired product 20 (40 mg, 99%). HR-EIMS: m/z [M+H]⁺409.1272 (calcd. 409.1287). ¹H NMR (400 MHz, DMSO-d₆) δ 7.48 (1H, d,J=2.1 Hz, H-2), 7.03 (1H, d, J=7.9 Hz, H-5′), 6.99 (1H, d, J=4.0 Hz,H-5), 6.89 (1H, dd, J=5.8, 1.5 Hz, H-2′), 6.76 (1H, ddd, J=7.8, 6.0, 1.7Hz, H-6′), 6.12 (2H, s, —OCH₂O—), 5.70 (1H, m, CH—OH), 5.59 (2H, br s,H-9), 3.97 (3H, s, 3-OCH₃), 3.64 (3H, s, 4-OCH₃), 1.51 (3H, dd, J=6.3,1.3 Hz, Me). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.5 (C-9′), 151.2 (C-3),149.2 (C-4), 147.0 (C-3′), 146.9 (C-4′), 137.2 (C-7), 136.2 (C-7′),134.7 (C-6), 128.8 (C-1), 128.6 (C-1′), 123.5 (C-8′), 123.4 (C-6′),118.2 (C-8), 110.9 (C-2′), 108.0 (C-5′), 105.8 (C-5), 102.9 (C-2), 101.2(—OCH₂O—), 68.9 (CH—OH), 65.6 (C-9), 55.8 (3-OCH₃), 55.1 (4-OCH₃), 23.3(CH₃).

Compound 21

To a solution of 19o (207 mg, 0.45 mmol) was added K₂CO₃. The mixturewas stirred for 2 h. TLC showed one spot with less polarity. The mixturewas purified by a silica gel column to give desired product 21 (122 mg,70%). HR-EIMS: m/z [M+H]⁺ 389.1000 (calcd. 389.1025). ¹H NMR (400 MHz,CDCl₃) δ 7.68 (1H, s, H-2), 7.14 (1H, s, H-5), 6.97 (1H, d, J=7.9 Hz,H-5′), 6.85 (1H, d, J=1.3 Hz, H-2′), 6.82 (1H, dd, J=7.9, 1.7 Hz, H-6′),6.03 (1H, br s, —OCH₂O—), 5.99 (1H, br s, —OCH₂O—), 5.43 (2H, s, H-9),4.10 (3H, s, 3-OCH₃), 3.82 (3H, s, 4-OCH₃), 3.78 (1H, s, C≡CH). ¹³C NMR(100 MHz, CDCl₃) δ 169.6 (C-9′), 152.7 (C-3), 150.3 (C-4), 147.8 (C-3′),147.6 (C-4′), 143.8 (C-7), 140.4 (C-7′), 133.3 (C-6), 128.8 (C-1′),127.9 (C-1), 123.5 (C-6′), 121.4 (C-8′), 118.4 (C-8), 110.5 (C-2′),108.3 (C-5′), 106.4 (C-5), 104.1 (C-2), 101.4 (—OCH₂O—), 87.7 (C≡CH),77.7 (C≡CH), 68.2 (C-9), 56.2 (3-OCH₃), 55.9 (4-OCH₃).

Compound 22

The crude ketimine 19p (115.8 mg, 0.21 mmol) was suspended in 1:1 (v/v)ratio of 1N HCl:THF (4 mL) solution and stirred at r.t. for 3-5 h. Uponconsumption of the ketimine as determined by TLC, THE was removed andthe aqueous residue was basified using 1N KOH to a pH of 10-14. Theaqueous layer was then extracted three times with ethyl acetate (30 mL).The organic layer was dried with anhydrous NaSO₄, filtered andconcentrated in vacuo to give the crude amine. Upon purification viacolumn chromatography the pure product 22 (70 mg, 84%) was obtainedafter solvent removal. HR-EIMS: m/z [M+H]⁺ 380.0849 (calcd. 380.1134).¹H NMR (400 MHz, DMSO-d₆) δ 7.62 (1H, s, H-2), 7.02 (1H d, J=7.9 Hz,H-5′), 6.95 (1H, s, H-5), 6.86 (1H, d, J=1.5 Hz, H-2′), 6.75 (1H, dd,J=7.9, 1.6 Hz, H-6′), 6.11 (2H, s, —OCH₂O—), 5.75 (1H, s, NH₂), 5.36(2H, s, H-9), 3.94 (3H, s, 3-OCH₃), 3.65 (3H, s, 4-OCH₃). ¹³C NMR (100MHz, DMSO-d₆) δ 169.9 (C-9′), 150.6 (C-3), 149.8 (C-4), 147.0 (C-3′),146.7 (C-4′), 145.1 (C-7), 134.8 (C-7′), 129.6 (C-6), 129.0 (C-1), 128.9(C-1′), 123.9 (C-6′), 123.5 (C-8′), 121.9 (C-8), 111.2 (C-2′), 108.0(C-5′), 105.5 (C-5), 101.2 (C-2), 101.0 (—OCH₂O—), 67.0 (C-9), 55.8(3-OCH₃), 55.3 (4-OCH₃).

Compound 23

To solution of 22 (38 mg, 0.1 mmol) and propynyl bromide (30 mg, 0.2mmol) in MeCN (2 mL) was added Cs₂CO₃. The mixture was stirred for 12 hat 60° C. After TLC show one spot with less polarity, the reactionsolution was quenched with water and extracted with CH₂Cl₂. The obtainedsolution was concentrated and purified by prep-TLC to give desiredproduct 23 (4.6 mg, 11%). HR-EIMS: m/z [M+H]⁺ 418.1279 (calcd.418.1291). ¹H NMR (400 MHz, CDCl₃) δ 7.19 (1H, d, J=4.2 Hz, H-2), 7.14(1H, s, H-5), 6.97 (1H, d, J=6.1 Hz, H-5′), 6.83 (1H, d, J=1.6 Hz,H-2′), 6.80 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.10 (2H, m, —OCH₂O—), 6.06(2H, m, —OCH₂O—), 5.75 (1H, s, NH), 5.30 (2H, s, H-9), 4.16 (2H, s,CH₂C≡CH), 4.06 (3H, s, 3-OCH₃), 3.83 (1H, s, CH₂C≡CH), 3.81 (3H, s,4-OCH₃). ¹³C NMR (100 MHz, DMSO-d₆) δ 169.7 (C-9′), 150.5 (C-3), 149.7(C-4), 146.8 (C-3′), 146.6 (C-4′), 144.9 (C-7), 134.8 (C-7′), 129.5(C-1), 129.5 (C-1′), 128.7 (C-6), 123.8 (C-6′), 123.3 (C-8′), 121.7(C-8), 111.0 (C-2′), 107.9 (C-5′), 105.4 (C-5), 101.0 (C-2), 100.7(—OCH₂O—), 79.7 (CH₂C≡CH), 74.1 (CH₂C≡CH), 66.6 (C-9), 55.6 (3-OCH₃),55.1 (4-OCH₃), 33.2 (CH₂C≡CH).

Compound 24

A solution of 22 (38 mg, 0.1 mmol) in dry pyridine (2 mL) was added Ac₂O(14.2 μL, 0.15 mmol). The resulting mixture was stirred under r.t.overnight. After the reaction was complete, it was diluted by CH₂Cl₂ andwashed with 10% HCl solution. The combined organic phase was furtherextracted with a saturate aqueous solution of NaHCO₃ and dried overNa₂SO₄. After concentration, the crude product was purified prep-TLC(1:1 n-hexane:ethyl acetate), to give 24 (15 mg, 36%) as a white solid.HR-EIMS: m/z [M+H]⁺ 433.1149 (calcd. 422.1240). ¹H NMR (400 MHz, CDCl₃)δ 7.18 (1H, s, H-2), 7.01 (1H, s, H-5), 6.82 (1H, d, J=7.9 Hz, H-5′),6.77 (1H, d, J=1.6 Hz, H-2′), 6.72 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.01(1H, d, J=1.4 Hz, —OCH₂O—), 5.93 (1H, d, J=1.5 Hz, —OCH₂O—), 5.17 (21-1,s, H-9), 3.90 (3H, s, 3-OCH₃), 3.75 (3H, s, 4-OCH₃), 2.30 (3H, d, J=0.9Hz, OAc). ¹³C NMR (100 MHz, CDCl₃) δ 170.6 (OAc), 169.7 (C-9′), 152.2(C-3), 150.1 (C-4), 147.6 (C-3′), 147.6 (C-4′), 138.5 (C-7), 135.7(C-7′), 130.0 (C-1), 128.9 (C-1′), 128.2 (C-6), 124.4 (C-8′), 123.6(C-6′), 118.6 (C-8), 110.7 (C-2′), 108.2 (C-5′), 106.5 (C-5), 101.4(C-2), 100.9 (—OCH₂O—), 68.3 (C-9), 56.1 (3-OCH₃), 55.9 (4-OCH₃), 23.40(OAc).

General Procedure A: Glycosylated Modification

A solution of sugar (0.61 mmol) in dry pyridine (2 mL) was added aceticanhydride (Ac₂O) (0.4 mL, 4.21 mmol). The resulting mixture was stirredunder r.t. overnight. After the reaction was complete, it was diluted bydichloromethane (CH₂Cl₂) and washed with 10% hydrogen chloride (HCl)solution. The combined organic phase was further extracted with asaturated aqueous solution of sodium bicarbonate (NaHCO₃) and dried oversodium sulfate (Na₂SO₄). After concentration, the crude product wasdirectly used for next step without further purification.

The crude per-acetylated glycoside was dissolved in CH₂Cl₂ (5 mL) andcooled to 0° C. before the drop-wise addition of hydrogen bromide (33%in acetic acid, 1.5 mL). The reaction was warmed to r.t. and stirred foran additional 4 h. Following completion, the reaction was quenched withwater, and the aqueous layer was extracted with CH₂Cl₂. The organiclayers were combined and washed with water, sodium bicarbonate, andbrine, dried with Na₂SO₄, and concentrated under reduced pressure toafford the glycosyl bromide. After concentration, the crude product wasdirectly used for next step without further purification.

Diphyllin (5) (215 mg, 0.61 mmol) was dissolved in chloroform (CHCl₃)(15 mL) and aqueous sodium hydroxide (NaOH) (0.1 M, 20 mL) prior toadding tetra-n-butylammonium bromide (TBAB) (306 mg, 0.95 mmol). Thereaction was then heated to 40° C. for 10 min before adding the glycosylbromide (0.61 mmol), and the reaction was then stirred at 40° C.overnight. After cooling to r.t., the aqueous layer was extracted withCHCl₃ and the combined organic layers were washed with brine, dried withNa₂SO₄ and concentrated under reduced pressure. Flash chromatographicseparation of the reaction mixture (silica gel, 45%-52% ethyl acetate inn-hexane) afford the target diphyllin per-acetylated glycoside(25a-25g). The reaction mixtures of compounds 25c-25g were not purified,but they were directly treated with basic reagents to produce theircorresponding hydrolyzed compounds 26c-26g.

Potassium carbonate (K₂CO₃) (168.4 mg, 1.2 mmol) was added to a solutionof a diphyllin per-acetylated glycoside (25a-25g) in MeOH (10 mL) andstirred at r.t. for 30 min. When no starting material was observed byTLC, the reaction was quenched by the addition of dilute HCl (2 M),filtered through filter paper, and concentrated under reduced pressure.Flash chromatographic separation of the reaction mixture (silica gel,2%-10% MeOH in CHCl₃) afforded the target glycoside (26a-26g).

Peracetylated 7β-D-xylosyloxydiphyllin (25a, total yield 80%)

HR-EIMS: m/z [M+H]⁺ 639.5780 (calcd. 639.1714). ¹H NMR (400 MHz, CDCl₃)δ 7.51 (1H, d, J=1.0 Hz, H-2), 7.07 (1H, s, H-5), 6.95 (1H, d, J=7.8 Hz,H-5′), 6.84-6.80 (1H, m, H-2′), 6.80-6.76 (1H, m, H-6′), 6.11-6.03 (2H,m, —OCH₂O—), 5.50-5.44 (1H, m, H-1″), 5.44-5.40 (2H, m, H-9), 5.31 (1H,br t, J=8.9 Hz, H-3″), 5.13 (1H, dd, J=8.8, 5.2 Hz, H-2″), 5.12 (1H, dd,J=8.3, 6.2 Hz, H-4″), 4.26 (1H, dd, J=11.8, 5.2 Hz, H-5″a), 4.16-4.10(1H, m, H-5″b), 4.07 (3H, s, 3-OCH₃), 3.80 (3H, s, 4-OCH₃), 2.12 (3H, s,—OAc), 2.10 (3H, s, —OAc), 2.04 (3H, s, —OAc).

Peracetylated Patentiflorin A (25b, total yield 71%)

HR-EIMS: m/z [M+H]⁺ 653.1744 (calcd. 653.1870). ¹H NMR (400 MHz, CDCl₃)δ 7.53 (1H, d, J=1.9 Hz, H-2), 7.07 (1H, s, H-5), 6.96 (1H, d, J=7.8 Hz,H-5′), 6.85-6.81 (1H, m, H-2′), 6.81-6.77 (1H, m, H-6′), 6.12-6.04 (2H,m, —OCH₂O—), 5.50-5.43 (2H, m, H-9), 5.39 (1H, dd, J=14.7, 2.4 Hz,H-2″), 5.27 (1H, t, J=9.6 Hz, H-3″), 5.12 (1H, d, J=7.9 Hz, H-1″), 5.00(1H, t, J=9.6 Hz, H-4″), 4.07 (3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃),3.61 (1H, ddd, J=9.7, 6.1, 1.2 Hz, H-5″), 2.12 (3H, s, —OAc), 2.06 (3H,s, —OAc), 2.06 (3H, s, —OAc), 1.28 (3H, dd, J=6.1, 1.4 Hz, H-6″).

7β-D-xylosyloxydiphyllin (26a, total yield 66%)

HR-EIMS: m/z [M+H]⁺ 513.1389 (calcd. 513.1397). ¹H NMR (400 MHz,methanol-d₆) δ 8.13 (1H, s, H-2), 7.09 (1H, d, J=1.8 Hz, H-5), 6.97 (1H,d, J=7.9 Hz, H-5′), 6.82 (1H, dd, J=3.1, 1.3 Hz, H-2′), 6.79 (1H, dt,J=7.9, 1.8 Hz, H-6′), 6.08-6.04 (2H, m, —OCH₂O—), 5.58 (11H, dd, J=15.2,1.6 Hz, H-9), 5.46 (1H, dd, J=15.2, 2.0 Hz, H-9), 4.03 (3H, s, 3-OCH₃),3.98 (1H, d, J=6.5 Hz, H-1″), 3.98-3.92 (1H, m, H-5″), 3.74 (3H, s,4-OCH₃), 3.64 (1H, m, H-2″), 3.51-3.44 (1H, m, H-4″), 3.16-3.11 (2H, m,H-3″, H-5″).

Patentiflorin A (26b, total yield 68%)

HR-EIMS: m/z [M+H]⁺ 527.1526 (calcd. 527.1553). ¹H NMR (400 MHz, CD₃OD)δ 7.99-7.92 (1H, m, H-2), 6.86 (1H, m, H-5), 6.82-6.77 (1H, m, H-5′),6.63 (1H, m, H-2′), 6.59 (1H, m, H-6′), 5.94 (1H, d, J=1.2 Hz, —OCH₂O—),5.93 (1H, dd, J=2.2, 1.3 Hz, —OCH₂O—), 5.50-5.27 (2H, m, H-9), 4.67 (1H,d, J=7.9 Hz, H-1″), 3.88 (3H, s, 3-OCH₃), 3.57 (3H, s, 4-OCH₃), 3.54(1H, m, H-2″), 3.33 (1H, t, J=9.1 Hz, H-3″), 3.05 (1H, t, J=9.2 Hz,H-4″), 3.170 (1H, m, H-5″), 1.21 (3H, m, H-6″).

7-O-α-L-arabinosyldiphyllin (26c, total yield 45%)

HR-EIMS: m/z [M+H]⁺ 513.1305 (calcd. 513.1397). ¹H NMR (400 MHz,acetone-d₆) δ 8.21 (1H, s, H-2), 7.07 (1H, d, J=2.2, H-5), 6.95 (1H, d,J=7.9 Hz, H-5′), 6.85 (1H, dd, J=5.8, 1.7 Hz, H-2′), 6.79 (1H, dt,J=7.9, 2.0 Hz, H-6′), 6.10-6.04 (2H, m, —OCH₂O—), 5.62-5.34 (3H, m, H-9,H-1″), 4.98 (1H, dt, J=4.2, 1.9 Hz, OH-3″), 4.87 (1H, dd, J=7.1, 2.0 Hz,H-1″), 4.24 (1H, t, J=5.9 Hz, OH-2″), 4.03 (I H, ddd, J=8.7, 7.0, 4.3Hz, H-2″), 3.96 (3H, s, 3-OCH₃), 3.98-3.94 (1H, m, H-5″), 3.91-3.84 (1H,m, H-4″), 3.70 (3H, s, 4-OCH₃), 3.68-3.72 (1H, m, H-3″), 3.57 (1H, d,J=12.4 Hz, H-5″).

7-O-β-L-fucopyranosyldiphyllin (26d, total yield 49%)

HR-EIMS: m/z [M+H]⁺ 527.1433 (calcd. 527.1553). ¹H NMR (400 MHz,DMSO-d₆) δ 8.20 d (1.9 Hz, H-2), 7.05 (1H, dd, J=7.9, 1.4 Hz, H-5′),6.99 (1H, d, J=2.1 Hz, H-5), 6.94 (1H, dd, J=11.1, 1.5 Hz, H-2′), 6.81(1H, ddd, J=7.8, 6.0, 1.6 Hz, H-6′), 6.13 (2H, s, —OCH₂O—), 5.59-5.44(3H, m, H-9, OH-2″), 4.89 (1H, dd, J=14.7, 3.1 Hz, OH-3″), 4.70 (1H, dd,J=7.7, 4.9 Hz, H-1″), 4.20 (1H, d, J=7.3 Hz, OH-4″), 3.95 (3H, s,3-OCH₃), 3.75 (dd, J=9.4, 7.7 Hz, 1H), 3.67 (3H, s, 4-OCH₃), 3.62 (1H,d, J=6.4 Hz, H-2″), 3.50-3.42 (3H, m, H-3″, H-4″, H-5″), 1.19 (3H, d,J=6.2 Hz, H-6″).

7-O-β-D-fucopyranosyldiphyllin (26e, total yield 67%)

HR-EIMS: m/z [M+H]⁺ 527.1550 (calcd. 527.1553). ¹H NMR (400 MHz,acetone-d₆) S 8.29-8.08 (1H, m, H-2), 7.12-7.01 (1H, m, H-5), 6.96 (1H,q, J=7.8 Hz, H-5′), 6.81 (2H, m, H-2′, H-6′), 6.08 (2H, t, J=7.5 Hz,—OCH₂O—), 5.62-5.48 (1H, m, H-9), 5.50-5.35 (1H, m, H-9), 5.30-5.15 (1H,m, OH-2″), 4.87 (1H, dt, J=11.7, 6.2 Hz, H-1″), 4.69 (1H, s, OH-4″),4.02-3.95 (2H, m, H-3″, H-4″), 3.79 (3H, d, J=7.4 Hz, 3-OCH₃), 3.76-3.72(1H, m, H-2″), 3.70 (4H, m, 4-OCH₃, H-5″), 1.29 (q, J=8.2, 7.7 Hz, 3H).

7-O-β-D-glucopyranosyldiphyllin (26f, total yield 48%)

HR-EIMS: m/z [M+H]⁺ 543.1500 (calcd. 543.1503). ¹H NMR (400 MHz,acetone-d₆) S 8.22 (1H, dq, J=9.0, 4.9 Hz, H-2), 7.05 (1H, m, H-5), 6.95(1H, tq, J=8.2, 4.0 Hz, H-5′), 6.89-6.74 (2H, m, H-2′, H-6′), 6.07 (2H,m, —OCH₂O—), 5.73 (1H, m, H-9), 5.48-5.36 (1H, m, H-9), 5.22 (1H, q,J=4.3 Hz, OH-3″), 4.91 (1H, tt, J=7.6, 3.9 Hz, OH-4″), 4.65 (1H, d,J=4.6 Hz, H-1″), 4.43 (1H, d, J=4.8 Hz, OH-6″), 3.97 (4H, m, 3-OCH₃,H-6″), 3.70 (5H, m, 4-OCH₃, H-6″, H-2″), 3.61-3.43 (2H, m, H-5″, H-3″),3.39 (1H, m, H-4″).

7-O-β-D-galactopyranosyldiphyllin (26g, total yield 64%)

HR-EIMS: m/z [M+H]⁺ 543.1370 (calcd. 543.1503). ¹H NMR (400 MHz,acetone-d₆) δ 8.22 (1H, d, J=1.0 Hz, H-2), 7.10 (1H, s, H-5), 6.98 (1H,dt, J=7.9, 0.6 Hz, H-5′), 6.91-6.85 (1H, m, H-2′), 6.85-6.79 (1H, m,H-6′), 6.13-6.06 (2H, m, —OCH₂O—), 5.84-5.35 (2H, m, H-9), 4.96-4.83(1H, m, H-1″), 4.34-4.19 (1H, m, OH), 4.08-4.01 (2H, m, H-2″, H-4″),4.00 (3H, s, 3-OCH₃), 3.97-3.82 (3H, m, H-5″, H-6″), 3.73 (3H, s,4-OCH₃), 3.54-3.69 (1H, m, H-3″).

General Procedure B: Selective Modification of 3″-Hydroxy Group

The glycoside 26a or 26b (0.3 mmol) was allowed to react with aceticanhydride (32 μL, 0.32 mmol) in dry acetonitrile (MeCN) (1.5 mL) at 40°C. for 12 h in the presence of tetrabutylammonium acetate (TBOAc) (27mg, 0.09 mmol). The solution was concentrated in vacuo to yield amixture containing 27aa-27af (the total mixture yield from 26a: 52%) ora mixture containing 27ba-27bf (the total mixture yield from 26b: 50%)as a pale-yellow solid.

The compound mixture containing 27aa-27af was purified by flash columnchromatography (petroleum ether/EtOAc=1.5/1) to afford the six singlecompounds 27aa-27af (27aa, 27ab, 27ac, 27ad, 27ae and 27af). Thecompound mixture containing 27ba-27bf was purified by flash columnchromatography (petroleum ether/EtOAc=1.5/1) to afford the six singlecompounds 27ba-27bf (27ba, 27bb, 27bc, 27bd, 27be and 27bf).

To the suspension of the 27ab or 27bb (0.036 mmol) in CH₂Cl₂ (3 mL) wasadded triethylamine (Et₃N) (50 L, 0.36 mmol) and allyl chloroformate(AllocOCl) (10.7 μL, 0.1 mmol) at 0° C., and the reaction was slowlywarmed to r.t. After 12 h, the reaction mixture was poured into H₂O andwas extracted with CH₂Cl₂. The organic layer was washed with brine,dried over Na₂SO₄, and evaporated under reduced pressure to give acompound mixture containing 28ab1-28ab3 (the total mixture yield from27ab: 45%) or a compound mixture containing 28bb1-28bb3 (the totalmixture yield from 27bb: 52%) as white solids. The compound mixturecontaining 28ab1-28ab3 was purified by flash column chromatography(petroleum ether/EtOAc=1/1) to afford the three single compounds28ab1-28ab3 (28ab1, 28ab2 and 28ab3). The compound mixture containing28bb1-28bb3 was purified by flash column chromatography (petroleumether/EtOAc=1/1) to afford the three single compounds 28bb1-28bb3(28bb1, 28bb2 and 28bb3).

To a solution of 28ab3 or 28bb3 (0.004 mmol) in 6 mL of MeOH/CH₂Cl₂(v/v=1/2) was added acetyl chloride (AcCl) (15 μL) dropwise at 0° C. Thereaction mixture was stirred for 72 h, and then quenched with Et₃N. Thevolatile was removed in vacuo. The resulting residue was purified bysilica gel column chromatography (petroleum ether/EtOAc 1/1) to afford29a or 29b (18.4 mg, 67%) as white solids.

29a or 29b (0.05 mmol), Et₃N (20.8 μL, 0.15 mmol) and4-dimethylaminopyridine (DMAP) (catalyzed amount) were dissolved in 2 mLof dry CH₂Cl₂. Selected acyl chloride (0.15 mmol) was added to themixture at 0° C. The reaction mixture was allowed to warm up to 25° C.and kept stirred at 25° C. for 2 h. The mixture was purified by prep-TLC(petroleum ether/EtOAc=1/1) to give desired products 30a-30k.

To a solution of 29a (20 mg, 0.036 mmol) in dry THF (2 mL) were addedsuccessively, freshly powdered KOH (3.6 mg, 0.0648 mmol), 18-crown-6(0.4 mg, 0.0014 mmol) and benzyl bromide (11 μL, 0.09 mmol). The mixturewas stirred at r.t. and the reaction is monitored by TLC; at the end ofthe reaction, the mixture was diluted with CH₂Cl₂ and washed severaltimes with water. The combined organic phase dried over Na₂SO₄ andconcentrated. The obtained residue was purified by s prep-TLC (petroleumether/EtOAc=1/1) to give desired products 30l.

To a solution of 30a-30l (0.045 mmol) in dry THF (4 ml) were addedtriphenylphosphine (PPh₃) (5.9 mg, 0.023 mmol), Et₃N (31.2 μL, 0.225),HCOOH (17 μL, 0.45 mmol) andtetrakis(triphenylphosphine)palladium(0)[Pd(PPh₃)₄] (5 mg, 0.0025 mmol)in sequence. The solution was stirred for 4 h at 55° C. under nitrogen,and concentrated in vacuo to give a mixture, which was chromatographedon prep-TLC (petroleum ether/EtOAc=1/1) to give desired products31a-31l.

4″-Acetyl-7-O-β-D-xylosyldiphyllin (27aa)

HR-EIMS: m/z [M+H]⁺ 555.1406 (calcd. 555.1503). ¹H NMR (400 MHz,Acetone-d₆) δ 7.63 (1H, d, J=1.0 Hz, H-2), 7.07 (1H, d, J=2.3 Hz, H-5),7.01-6.94 (1H, m, H-5′), 6.85 (1H, d, J=1.6 Hz, H-2′), 6.80 (1H, ddd,J=7.9, 1.6, 0.4 Hz, H-6′), 6.10 (2H, dt, J=4.9, 1.0 Hz, —OCH₂O—),5.60-5.45 (2H, m, H-9), 5.33 (1H, d, J=7.8 Hz, H-1″), 5.22 (1H, dd,J=9.1, 7.8 Hz, H-2″), 4.08 (1H, dd, J=11.4, 5.2 Hz, H-5″a), 4.03 (3H, s,3-OCH₃), 3.70-3.80 (1H, m H-3″), 3.70-3.80 (1H, m H-4″), 3.73 (3H, s,4-OCH₃), 3.46 (1H, dd, J=11.3, 10.1 Hz, H-5″b), 2.09 (3H, s, OAc).

3″-Acetyl-7-O-β-D-xylosyldiphyllin (27ab)

HR-EIMS: m/z [M+H]⁺ 555.1439 (calcd. 555.1503). ¹H NMR (400 MHz,acetone-d₆) δ 8.08 (1H, d, J=3.3 Hz, H-2), 7.05 (1H, br s, H-5),6.99-6.93 (1H, m, H-5′), 6.89 (1H, m, H-2′), 6.79 (1H dd, J=1.8, 6.0 Hz,H-6′), 6.13-6.07 (2H, m, —OCH₂O—), 5.53 (1H, d, J=15.0 Hz, H-1″), 5.41(1H d, J=15.1 Hz, H-3″), 5.11-5.01 (2H, m, br s, H-9), 4.04 (1H, m,H-5″), 3.94 (3H, s, 3-OCH₃), 3.91-3.82 (1H, m, H-2″), 3.69 (3H, d, J=6.5Hz, 4-OCH₃), 3.47-3.35 (1H, m, H-5″), 2.10 (3H, s, OAc). ¹³C NMR (100MHz, acetone-d₆) δ 171.0 (OAc), 169.8 (C-9′), 153.0 (C-3), 151.5 (C-4),148.4 (C-3′), 148.3 (C-4′), 145.8 (C-7), 136.7 (C-7′), 131.4 (C-1),129.7 (C-6), 128.1 (C-1′), 124.6 (C-6′), 120.0 (C-8), 111.8 (C-8′),111.7 (C-2′), 108.7 (C-5′), 106.7 (C-5), 106.6 (C-1″), 102.4 (C-2),102.2 (—OCH₂O—), 78.8 (C-3″), 73.2 (C-2″), 69.1 (C-4″), 67.8 (C-9), 66.8(C-5″), 56.4 (3-OCH₃), 55.8 (4-OCH₃), 21.2 (—OAc).

3″, 4″-Diacetyl-7-O-β-D-xylosyldiphyllin (27ad)

HR-EIMS: m/z [M+H]⁺ 597.1533 (calcd. 597.1608). ¹H NMR (400 MHz,Acetone-d₆) δ 7.53 (1H, d, J=0.8 Hz, H-2), 7.02 (1H, d, J=2.1 Hz, H-5),6.91 (1H, d, J=7.9 Hz, H-5′), 6.79 (1H, dd, J=3.3, 1.4 Hz, H-2′), 6.74(1H, ddd, J=7.9, 2.7, 1.7 Hz, H-6′), 6.03 (2H, dt, J=4.8, 0.9 Hz,—OCH₂O—), 5.59-5.45 (2H, m, H-9), 5.42 (1H, d, J=7.6 Hz, H-1″), 5.23(1H, dd, J=9.8, 7.6 Hz, H-2″), 5.14 (1H, br t, J=9.3 Hz, H-3″), 4.06(1H, dd, J=11.5, 5.6 Hz, H-5″a), 3.95 (3H, s, 3-0 CH₃), 3.94 (1H, br td,J=10.3, 5.3 Hz, H-4″), 3.66 (3H, s, 4-OCH₃), 3.51 (1H, br t, J=10.8 Hz,H-5″b), 2.03 (3H, s, —OAc), 1.98 (3H, s, —OAc).

2″,4″-Diacetyl-7-O-β-D-xylosyldiphyllin (27ae)

HR-EIMS: m/z [M+H]⁺ 597.1509 (calcd. 597.1608). ¹H NMR (400 MHz,Acetone-d₆) δ 7.62 (1H, d, J=1.8 Hz, H-2), 7.03 (1H, d, J=2.1 Hz, H-5),6.92 (1H, dd, J=7.9, 1.1 Hz, H-5′), 6.80 (1H, d, J=1.6 Hz, H-2′),6.77-6.71 (1H, m, H-6′), 6.07-6.00 (2H, m, —OCH₂O—), 5.54-5.40 (2H, m,H-9), 5.43 (1H, d, J=7.1, H-1″), 5.27 (1H, dd, J=8.8, 7.2 Hz, H-2″),4.90 (1H, br td, J=9.0, 5.2 Hz, H-4″), 4.15 (1H, dd, J=11.6, 5.1 Hz,H-5″a), 4.04-3.99 (1H, br t, J=8.7 Hz, H-3″), 3.98 (3H, s, 3-OCH₃), 3.67(3H, s, 4-OCH₃), 3.50 (1H, dd, J=11.6, 9.5 Hz, H-5″b), 2.10 (3H, s,—OAc), 2.03 (3H, s, —OAc).

2″,3″-Diacetyl-7-O-β-D-xylosyldiphyllin (27af)

HR-EIMS: m/z [M+H]) 597.1505 (calcd. 597.1608). ¹H NMR (400 MHz,Acetone-d₆) δ 8.10 (1H, d, J=1.5 Hz, H-2), 7.10 (1H, d, J=4.5 Hz, H-5),6.98 (1H, dd, J=7.9, 1.2 Hz, H-5′), 6.88 (1H, br d, J=1.3 Hz, H-2′),6.81 (1H, dd, J=7.9, 1.7 Hz, H-6′), 6.10 (2H, ddd, J=4.8, 1.6, 1.1 Hz,—OCH₂O—), 5.58-5.41 (2H, m, H-9), 5.25 (1H, br t, J=9.5 Hz, H-3″), 5.13(1H, d, J=7.6 Hz, H-1″), 5.00 (1H, br td, J=9.6, 5.5 Hz, H-4″), 4.16(1H, dd, J=11.5, 5.5 Hz, H-5″a), 4.02 (1H, d, J=9.6, 7.7 Hz, H-2″), 3.97(3H, s, 3-OCH₃), 3.72 (3H, s, 4-OCH₃), 3.54 (1H, dd, J=11.5, 9.9 Hz,H-5″b), 2.06 (3H, s, —OAc), 1.98 (3H, s, —OAc).

4″-Acetyl-7-O-β-D-patentiflorin A (27ba)

HR-EIMS: m/z [M+H]⁺ 569.1612 (calcd. 569.1659). ¹H NMR (400 MHz, CDCl₃)δ 7.58 (1H, d, J=1.5 Hz, H-2), 7.07 (1H, s, H-5), 6.96 (1H, d, J=7.8 Hz,H-5′), 6.86-6.81 (1H, m, H-2′), 6.81-6.77 (1H, m, H-6′), 6.12-6.01 (2H,n, —OCH₂O—), 5.51-5.37 (2H, m, H-9), 5.272 (1H, dd, J=9.8, 7.9 Hz,H-2″), 5.06 (1H, d, J=7.8 Hz, H-1″), 4.07 (3H, s, 3-OCH₃), 3.81 (3H, s,4-OCH₃), 3.67 (1H, t, J=9.2 Hz, H-3″), 3.44 (1H, m, H-4″), 3.43 (1H, m,H-5″), 2.20 (3H, s, OAc), 1.39 (3H, d, J=4.0 Hz, H-6″).

3″-Acetyl-7-O-β-D-patentiflorin A (27bb)

HR-EIMS: m/z [M+H]⁺ 569.1602 (calcd. 569.1659). ¹H NMR (400 MHz, CDCl₃)δ 7.93 (1H, s, H-2), 7.02 (1H, d, J=3.6 Hz, H-5), 6.91 (1H, dd, J=9.2,8.0 Hz, H-5′), 6.82-6.67 (2H, m, H-2′,6′), 6.11-5.99 (2H, m, —OCH₂O—),5.55-5.34 (2H, m, H-9), 4.86 (1H, t, J=9.2 Hz, H-3″), 4.82 (1H, d, J=7.8Hz, H-1″), 3.99 (3H, s, 3-OCH₃), 3.93 (1H, dd, J=9.5, 7.8 Hz, H-2″),3.77 (3H, d, J=2.3 Hz, 4-OCH₃), 3.42 (1H, t, J=9.2 Hz, H-4″), 3.37-3.28(1H, m, H-5″), 2.20 (3H, s, —OAc), 1.34 (3H, dd, J=6.0, 1.2 Hz, H-6″).¹³C NMR (100 MHz, CDCl₃) δ 173.5 (OAc), 170.3 (C-9′), 152.1 (C-3), 150.3(C-4), 147.6 (C-3′), 147.6 (C-4′), 144.5 (C-7), 136.8 (C-7′), 131.2(C-8), 130.8 (C-6), 128.3 (C-1′), 127.4 (C-1), 123.7 (C-6′), 119.1(C-8′), 110.8 (C-2′), 108.3 (C-5′), 106.2 (C-1″), 104.7 (C-5), 101.4(C-2), 101.1 (—OCH₂O—), 79.4 (C-3″), 73.9 (C-4″), 73.0 (C-2″), 72.4(C-5″), 67.7 (C-9), 56.4 (3-OCH₃), 55.9 (4-OCH₃), 21.20 (C-6″), 17.7(OAc).

2″-Acetyl-7-O-β-D-patentiflorin A (27bc)

HR-EIMS: m/z [M+H]⁺ 569.1596 (calcd. 569.1659). ¹H NMR (400 MHz, CDCl₃)δ 7.96 (1H, s, H-2), 7.09 (1H, s, H-5), 6.96 (1H, d, J=7.9 Hz, H-5′),6.84 (1H, dd, J=3.1, 1.4 Hz, H-2′), 6.83-6.78 (1H, m, H-6′), 6.12-6.03(2H, m, —OCH₂O—), 5.54-5.39 (2H, m, H-9), 4.83 (1H, dt, J=8.7, 1.4 Hz,H-4″), 4.74 (1H, t, J=9.5 Hz, H-1″), 4.04 (3H, s, 3-OCH₃), 3.92 (1H, dd,J=9.3, 7.9 Hz, H-2″), 3.81 (3H, s, 4-OCH₃), 3.67 (1H, t, J=9.3 Hz,H-3″), 3.51-3.44 (1H, m, H-5″), 2.15 (3H, s, OAc), 1.27 (3H, d, J=1.5Hz, H-6″).

3″, 4″-Diacetyl-7-O-β-D-patentiflorin A (27bd)

HR-EIMS: m/z [M+H]⁺ 611.1705 (calcd. 609.1972). ¹H NMR (400 MHz, CDCl₃)δ 7.53 (1H, d, J=1.5 Hz, H-2), 7.07 (1H, s, H-5), 6.98-6.94 (1H, m,H-5′), 6.84-6.80 (1H, m, H-2′), 6.81-6.76 (1H, m, H-6′), 6.11-6.02 (2H,m, —OCH₂O—), 5.50-5.43 (2H, m, H-9), 5.42-5.36 (1H, m, H-2″), 5.12 (1H,d, J=7.9 Hz, H-1″), 5.06 (1H, t, J=9.8 Hz, H-3″), 4.06 (3H, s, 3-OCH₃),3.80 (3H, s, 4-OCH₃), 3.54 (1H, t, J=9.1 Hz, H-4″), 3.48 (1H, ddd,J=8.8, 5.6, 1.1 Hz, H-5″), 2.14 (3H, s, —OAc), 2.11 (3H, s, —OAc), 1.40(3H, dd, J=5.8, 1.2 Hz, H-6″).

2″,4″-Diacetyl-7-O-β-D-patentiflorin A (27be)

HR-EIMS: m/z [M+H]⁺ 611.1721 (calcd. 609.1972). ¹H NMR (400 MHz, CDCl₃)δ 7.58 (1H, d, J=1.7 Hz, H-2), 7.07 (1H, s, H-5), 6.96 (1H, d, J=7.9 Hz,H-5′), 6.83 (1H, dd, J=3.0, 1.5 Hz, H-2′), 6.80 (1H, ddd, J=7.9, 3.0,1.7 Hz, H-6′), 6.12-6.03 (2H, m, —OCH₂O—), 5.50-5.38 (2H, m, H-9),5.38-5.29 (1H, m, H-2″), 5.05 (1H, d, J=7.9 Hz, H-1″), 4.84 (1H, t,J=9.5 Hz, H-4″), 4.07 (3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃), 3.77 (1H,t, J=9.6 Hz, H-3″), 3.60-3.50 (1H, m, H-5″), 2.20 (3H, s, —OAc), 2.15(3H, s, —OAc), 1.29 (3H, dd, J=6.2, 1.4 Hz, H-6″).

2″,3″-Diacetyl-7-O-β-D-patentiflorin A (27bf)

HR-EIMS: m/z [M+H]⁺ 611.1699 (calcd. 609.1972). ¹H NMR (400 MHz,Chloroform-d) δ 7.93 (1H, s, H-2), 7.08 (1H, s, H-5), 6.97-6.93 (1H, m,H-5′), 6.86-6.81 (1H, m, H-2′), 6.80-6.76 (1H, m, H-6′), 6.11-6.02 (2H,m, —OCH₂O—), 5.53-5.37 (2H, m, H-9), 5.08 (1H, t, J=9.5 Hz, H-3′), 4.92(1H, t, J=9.6 Hz, H-4″), 4.88 (1H, d, J=7.8 Hz, H-1″), 4.04 (1H, dd,J=9.3, 7.9 Hz, H-2″), 4.02 (3H, s, 3-OCH₃), 3.80 (3H, s, 4-OCH₃),3.55-3.46 (1H, m, H-5″), 2.15 (3H, s, —OAc), 2.06 (3H, s, —OAc), 1.24(3H, d, J=5.6 Hz, H-6″).

4″-O-allyl oxycarbonyl-3″-acetyl-7-O-β-D-xylosyldiphyllin (28ab1)

HR-EIMS: m/z [M+H]⁺ 639.1514 (calcd. 639.1714). ¹H NMR (400 MHz,Acetone-d₆) δ 7.66 (1H, s, H-2), 7.10 (1H, d, J=3.3 Hz, H-5), 6.98 (1H,dd, J=7.9, 1.8 Hz, H-5′), 6.92-6.77 (2H, m, H-2′, H-6′), 6.10 (2H, dt,J=4.8, 1.3 Hz, —OCH₂O—), 5.96 (1H, ddt, J=16.2, 10.9, 5.6 Hz, O-Alloc),5.63-5.46 (2H, m, H-9), 5.44 (1H, d, J=7.6 Hz, H-1″), 5.34 (1H, dq,J=17.2, 1.6 Hz, O-Alloc), 5.24 (1H, bt t, J=9.4 Hz, H-3″) 5.29-5.20 (1H,m, O-Alloc), 5.08 (1H, dd, J=9.8, 7.8 Hz, H-2″), 4.85 (1H, dd, J=5.1,2.1 Hz, O-Alloc), 4.71 (1H, m, O-Alloc), 4.10 (1H, dd, J=11.2, 5.5 Hz,H-5″a), 4.02 (3H, s, 3-OCH₃), 3.99 (1H, br td, J=10.2, 5.5 Hz, H-4″),3.72 (3H, s, 4-OCH₃), 3.51 (1H, br t, J=10.9 Hz, H-5″b), 2.08 (3H, s,OAc).

2″-O-allyl oxycarbonyl-3″-acetyl-7-O-β-D-xylosyldiphyllin (28ab2)

HR-EIMS: m/z [M+H]⁺ 639.1514 (calcd. 639.1714). ¹H NMR (400 MHz,Acetone-d₆) δ 8.09 (1H, s, H-2), 7.10 (1H, dd, J=4.1, 1.6 Hz, H-5), 6.98(1H, dd, J=7.9, 1.6 Hz, H-5′), 6.91-6.85 (1H, m, H-2′), 6.85-6.78 (1H,m, H-6′), 6.10 (2H, dt, J=5.2, 1.3 Hz, —OCH₂O—), 5.93 (1H, ddt, J=16.2,10.8, 5.6 Hz, O-Alloc), 5.76 (1H, dt, J=5.0, 2.4 Hz, O-Alloc), 5.60-5.41(2H, m, H-9), 5.29 (1H, bt t, J=9.5 Hz, H-3″), 5.25-5.20 (1H, m,O-Alloc), 5.14 (1H, d, J=7.6 Hz, H-1″), 4.88 (1H, br td, J=9.5, 5.5 Hz,H-4″), 4.68-4.54 (2H, m, O-Alloc), 4.26 (1H, dd, J=11.6, 5.5 Hz, H-5″a),4.03 (1H, dd, J=9.5, 7.6 Hz, H-2″), 3.96 (3H, s, 3-OCH₃), 3.72 (3H, s,4-OCH₃), 3.61 (1H, br t, J=11.2 Hz, H-5″b), 2.06 (3H, s, OAc).

2″,4″-O-diallyl oxycarbonyl-3″-acetyl-7-O-β-D-xylosyldiphyllin (28ab3)

HR-EIMS: m/z [M+H]⁺ 723.1703 (calcd. 723.1925). ¹H NMR (400 MHz, CDCl₃)δ 7.55 (1H, s, H-2), 7.09 (1H, s, H-5), 6.96 (1H, dd, J=8.2, 1.0 Hz,H-2′), 6.87-6.77 (2H, m, H-5′,6′), 6.12-6.04 (2H, m, —OCH₂O—), 5.90 (2H,m, H-9), 5.52-5.20 (8H, m, O-alloc), 5.05 (1H, d, J=7.7 Hz, H-1″),5.04-4.95 (1H, m, H-3″), 4.75-4.59 (4H, m, O-alloc), 4.24 (1H, dd,J=11.8, 5.5 Hz, H-5″), 4.06 (3H, s, 3-OCH₃), 3.81 (3H, s, 4-OCH₃), 3.36(1H, dd, J=11.7, 10.1 Hz, H-5″), 2.11 (3H, s, OAc). ¹³C NMR (100 MHz,acetone-d₆) δ 170.2 (OAc), 169.6 (C-9′), 155.3 (O-alloc), 155.1(O-alloc), 153.2 (C-3), 151.5 (C-4), 148.4 (C-3′), 148.3 (C-4′), 144.7(C-7), 132.9 (O-alloc), 132.8 (O-alloc), 136.7 (C-7′), 131.4 (C-1),129.6 (C-6), 127.2 (C-1′), 124.6 (C-6′), 120.1 (C-8), 119.2 (O-alloc),118.9 (O-alloc), 111.7 (C-2′), 111.7 (C-8′), 108.7 (C-1″, C-5′), 106.9(C-5), 102.5 (C-2), 102.2 (—OCH₂O—), 79.1 (C-3″), 75.7 (C-2″), 72.1(C-4″), 69.6 (O-alloc), 69.3 (Alloc), 67.5 (C-9), 63.4 (C-5″), 56.5(3-OCH₃), 55.8 (4-OCH₃), 21.2 (O-alloc).

2″,4″-O-diallyl oxycarbonyl-3″-acetyl-7-O-β-D-patentiflorin A (28bb3)

HR-EIMS: m/z [M+H]⁺ 737.1950 (calcd. 737.2082). ¹H NMR (400 MHz, CDCl₃)δ 7.56 (1H, s, H-2), 7.08 (1H, s, H-5), 7.00-6.91 (1H, m, H-5′),6.86-6.73 (2H, m, H-2′,6′), 6.08 (1H, br s, OCH₂O) 6.04 (I H, br s,—OCH₂O—), 5.90 (2H, m, H-9), 5.53-5.20 (9H, m, O-alloc, 2″,4″), 5.08(1H, dd, J=8.5, 4.4 Hz, H-1″), 4.78 (1H, td, J=9.4, 4.5 Hz, H-3″), 4.65(4H, dt, J=22.0, 5.1 Hz, O-alloc), 4.04 (3H, s, 3-OCH₃), 3.79 (3H, s,4-OCH₃), 3.58 (1H, dq, J=10.5, 5.0 Hz, H-5″), 2.06 (3H, s, OAc), 1.27(3H, d, J=25.1 Hz, H-6″). ¹³C NMR (100 MHz, CDCl₃) δ 170.1 (OAc), 169.6(C-9′), 154.1 (O-alloc), 152.2 (C-3), 150.3 (C-4), 147.6 (C-3′), 147.5(C-4′), 143.3 (C-7), 137.0 (C-7′), 131.5 (C-6), 131.2 (C-8), 131.0(C-1), 130.8 (O-alloc), 128.1 (C-1′), 123.6 (C-6′), 119.9 (O-alloc),119.4 (O-alloc), 118.9 (C-8′), 110.6 (C-2′), 108.2 (C-5′), 106.3 (C-1″),101.5 (C-5), 101.3 (C-2), 100.0 (—OCH₂O—), 75.9 (C-3″), 72.3 (C-2″),70.4 (C-4″), 69.4 (O-alloc), 69.1 (O-alloc), 68.5 (C-5″), 67.2 (C-9),56.6 (3-OCH₃), 55.9 (4-OCH₃), 20.7 (C-6″), 17.4 (OAc).

2″,4″-O-diallyl oxycarbonyl-7-O-β-D-xylosyldiphyllin (29a, yield 92%)

HR-EIMS: m/z [M+H]⁺ 681.1696 (calcd. 681.1819). H NMR (400 MHz,acetone-d₆) δ 7.58 (1H, s, H-2), 6.99 (1H, s, H-5), 6.86 (1H, m, H-5′),6.72 (2H, m, H-2′,6′), 5.98 (2H, m, H-9), 5.85 (9H, m, O-alloc, —OCH₂O—,H-1″, H-2″, H-4″), 4.73 (1H, d, J=7.7 Hz, H-5″), 4.56 (4H, m, O-alloc),4.12 (1H, m, H-3″), 3.92 (3H, s, 3-OCH₃), 3.61 (3H, s, 4-OCH₃), 3.42(1H, m, H-5″). ¹³C NMR (100 MHz, acetone-d₆) δ 169.6 (C-9′), 155.4(O-alloc), 155.2 (O-alloc), 153.2 (C-3), 151.6 (C-4), 148.4 (C-3′),148.3 (C-4′), 144.7 (C-7), 138.1 (C-7′), 132.9 (O-alloc), 132.8(O-alloc), 131.4 (C-1), 129.1 (C-6), 127.2 (C-1′), 124.6 (C-8), 124.5(C-6′), 120.1 (O-alloc), 119.2 (O-alloc), 118.9 (C-8′), 111.7 (C-2′),108.7 (C-1″, C-5′), 106.9 (C-5), 102.6 (C-2), 102.2 (—OCH₂O—), 78.7(C-3″), 75.7 (C-2″), 72.1 (C-4″), 69.6 (O-alloc), 69.3 (O-alloc), 67.5(C-9), 63.4 (C-5″), 56.5 (3-OCH₃), 55.8 (4-OCH₃).

2″,4″-O-diallyl oxycarbonyl-7-O-β-D-patentiflorin A (29b, yield 83%)

HR-EIMS: m/z [M+H]⁺ 695.1911 (calcd. 695.1976). ¹H NMR (400 MHz, CDCl₃)δ 7.62 (11H, s, H-2), 7.09 (1H, s, H-5), 6.95 (1H, dd, J=8.2, 1.4 Hz,H-5′), 6.84-6.77 (2H, m, H-2′,6′), 6.12-6.01 (2H, m, —OCH₂O—), 5.93 (2H,m, O-alloc), 5.50-5.25 (6H, m, H-9, O-alloc), 5.15 (1H, dd, J=9.7, 8.1Hz, H-1″), 4.99 (1H, d, J=8.1 Hz, H-2″), 4.74-4.63 (5H, m, H-4″,O-alloc), 4.06 (3H, s, 3-OCH₃), 3.88 (1H, t, J=9.5 Hz, H-3″), 3.80 (31H,s, 4-OCH₃), 3.49 (1H, ddt, J=12.7, 6.6, 4.9 Hz, H-5″), 1.31 (3H, dd,J=6.1, 1.7 Hz, H-6″). ¹³C NMR (100 MHz, CDCl₃) δ 169.8 (C-9′), 155.0(O-alloc), 154.9 (O-alloc), 152.3 (C-3), 150.4 (C-4), 147.7 (C-3′),147.7 (C-4′), 143.7 (C-7), 137.0 (C-7′), 131.1 (C-8), 130.9 (C-1), 128.3(C-6), 126.9 (C-1′), 123.7 (C-6′), 120.2 (O-alloc), 119.7 (O-alloc),119.3 (C-8′), 110.8 (C-2′), 108.3 (C-5′), 106.4 (C-1″), 101.6 (C-5),101.4 (C-2), 100.2 (—OCH₂O—), 79.3 (C-3″), 78.2 (C-1″), 73.5 (C-4″),70.5 (C-5″), 69.7 (O-alloc), 69.4 (O-alloc), 67.3 (C-9), 56.7 (3-OCH₃),56.0 (4-OCH₃), 17.5 (C-6″).

2″,4″-O-diallyloxycarbonyl-3″-O-dimethylcarbamyl-7-O-β-D-xylosyldiphyllin (30a, yield99%)

HR-EIMS: m/z [M+H]⁺ 752.2133 (calcd. 752.2191). ¹H NMR (400 MHz, CDCl₃)δ 7.56 (1H, s, H-2), 7.07 (1H, d, J=1.8 Hz, H-5), 6.93 (1H, dd, J=8.1,1.1 Hz, H-5′), 6.83-6.74 (2H, m, H-2′, H-6′), 6.05 (2H, dt, J=17.9, 1.5Hz, —OCH₂O—), 5.89 (2H, ddtd, J=17.6, 10.4, 5.8, 1.5 Hz, O-Alloc),5.52-5.39 (2H, m, H-9), 5.35 (1H, m, H-1″), 5.31 (1H, br t, J=1.6 Hz,H-3″), 5.27-5.23 (4H, m, O-Alloc), 5.10-5.06 (1H, m, H-2″), 5.02 (1H,tdd, J=9.7, 4.8, 2.1 Hz, H-4″), 4.73-4.59 (4H, m, O-Alloc), 4.20 (1H,ddd, J=11.8, 5.4, 1.1 Hz, H-5″a), 4.04 (3H, s, 3-OCH₃), 3.78 (3H, s,4-OCH₃), 3.40 (1H, ddd, J=11.9, 9.9, 0.9 Hz, H-5″b), 2.91 [6H, s,OC(O)N(CH₃)₂].

2″,4″-O-diallyloxycarbonyl-3″-O-(3,5-dinitrobenzoyl)-7-O-β-D-xylosyldiphyllin (30b,yield 90%)

HR-EIMS: m/z [M+H]⁺ 889.1763 (calcd. 889.1940). ¹H NMR (400 MHz, CDCl₃)δ 9.26 (1H, t, J=2.1 Hz, 3,5-dinitrobenzoyl), 9.15 (2H, d, J=2.1 Hz,3,5-dinitrobenzoyl), 7.59 (1H, s, H-2), 7.09 (1H, d, J=1.2 Hz, H-5),6.94 (1H, dd, J=7.9, 3.8 Hz, H-5′), 6.81 (1H, dd, J=9.7, 1.6 Hz, H-2′),6.78-6.72 (1H, m, H-6′), 6.10-6.01 (2H, m, —OCH₂O—), 5.76 (2H, ddt,J=17.2, 16.1, 5.7 Hz, O-Alloc), 5.61 (1H, t, J=9.4 Hz, O-Alloc),5.54-5.48 (1H, m, O-Alloc), 5.48-5.43 (2H, m, H-9), 5.24 (1H, d, J=8.2Hz, H-1″), 5.20 (2H, ddd, J=8.0, 2.8, 1.4 Hz, O-Alloc), 5.14 (1H, dd,J=6.4, 1.2 Hz, H-2″), 5.11 (1H, dd, J=6.5, 1.2 Hz, H-3″), 5.02 (1H, t,J=9.5 Hz, H-4″), 4.55 (4H, ddt, J=20.0, 5.7, 1.4 Hz, O-Alloc), 4.06 (3H,s, 3-OCH₃), 3.79 (3H, s, 4-OCH₃), 3.73-3.67 (1H, m, H-5″), 1.37 (3H, dd,J=6.1, 1.7 Hz, H-6″).

3″-O-dimethylcarbamyl-7-O-β-D-xylosyldiphyllin (31a, yield 69% of twosteps)

HR-EIMS: m/z [M+H]⁺ 584.1723 (calcd. 584.1768). ¹H NMR (400 MHz,acetone-d₆) δ 8.15 (1H, d, J=1.6 Hz, H-2), 7.10 (1H, d, J=4.4 Hz, H-5),6.98 (1H, dd, J=7.9, 1.5 Hz, H-5′), 6.88 (1H, dd, J=15.7, 1.6 Hz, H-2′),6.83 (1H, td, J=7.9, 1.7 Hz, H-6′), 6.10 (2H, dt, J=4.8, 1.3 Hz,—OCH₂O—), 5.56-5.38 (3H, m, H-9, H-1″), 5.02 (1H, d, J=7.6 Hz, H-3″),4.79-4.67 (2H, m, OH-2″, OH-4″), 4.05 (1H, ddd, J=11.5, 5.5, 1.9 Hz,H-2″), 3.97 (3H, s, 3-OCH₃), 3.89 (2H, m, H-4″, H-5″), 3.73 (3H, d,J=1.4 Hz, 4-OCH₃), 3.39 (1H, ddd, J=11.4, 10.0, 1.1 Hz, H-5″), 3.02 (3H,s, OCON(CH₃)₂), 2.93 (3H, s, OCON(CH₃)₂). ¹³C NMR (100 MHz, acetone-d₆)δ 169.8 (C-9′), 158.3 (OCON(CH₃)₂), 153.0 (C-3), 151.5 (C-4), 148.4(C-3′), 148.3 (C-4′), 145.9 (C-7), 136.7 (C-7′), 131.5 (C-1), 131.4(C-6), 129.7 (C-1′), 128.1 (C-8), 124.6 (C-6′), 120.0 (C-8′), 111.8(C-2′), 111.7 (C-5′), 108.7 (C-1″), 106.8 (C-2), 102.4 (C-5), 102.2(—OCH₂O—), 81.5 (C-3″), 73.5 (C-2″), 69.3 (C-4″), 67.8 (C-9), 66.7(C-5″), 56.3 (3-OCH₃), 55.8 (4-OCH₃), 36.7 (OCON(CH₃)₂), 36.2(OCON(CH₃)₂).

3″-O-(3,5-dinitrobenzoyl)-7-O-β-D-patentiflorin A (31b, yield 77% of twosteps)

HR-EIMS: m/z [M+H]⁺ 721.1337 (calcd. 721.1517). ¹H NMR (400 MHz,acetone-d₆) δ 9.17 (3H, m, 3,5-Dinitrobenzoyl), 8.11 (1H, dt, J=6.9, 3.1Hz, H-2), 7.13-7.05 (1H, m, H-5), 6.99 (1H, dd, J=7.7, 1.7 Hz, H-5′),6.89 (1H, dd, J=11.2, 1.5 Hz, H-2′), 6.83 (1H, ddd, J=9.4, 5.5, 1.6 Hz,H-6′), 6.16-6.08 (2H, m, —OCH₂O—), 5.75-5.41 (4H, m, H-9, OH-2″, OH-4″),5.18 (1H, d, J=7.8 Hz, H-1″), 4.95 (1H, d, J=4.9 Hz, H-2″), 4.21 (1H,ddd, J=9.7, 7.8, 5.3 Hz, H-4″), 3.95 (3H, s, 3-OCH₃), 3.71 (5H, m,4-OCH₃, H-3″, H-5″), 1.41 (3H, d, J=5.1 Hz, H-6″). ¹³C NMR (100 MHz,acetone-d₆) δ 169.8 (C-9′), 163.4 (3,5-dinitrobenzoyl), 152.9 (C-3),151.4 (3,5-dinitrobenzoyl), 151.3 (3,5-dinitrobenzoyl), 149.7 (C-4),148.3 (C-3′), 148.3 (C-4′), 145.8 (C-7), 136.7 (C-7′), 134.8(3,5-dinitrobenzoyl), 131.4 (C-1), 130.1 (3,5-dinitrobenzoyl), 129.6(C-8), 128.1 (C-6), 124.6 (C-1′), 123.3 (C-6′), 120.0(3,5-dinitrobenzoyl), 111.7 (C-8′), 111.7 (C-2′), 108.7 (C-5′), 106.7(C-2), 105.5 (C-1″), 102.3 (C-5), 102.2 (—OCH₂O—), 81.4 (C-3″), 74.1(C-2″), 73.4 (C-4″), 73.0 (C-5″), 67.8 (C-9), 56.3 (3-OCH₃), 55.8(4-OCH₃), 18.1 (C-6″).

3″-O-butyryl-7-O-β-D-patentiflorin A (31c, yield 77% of two steps)

HR-EIMS: m/z [M+H]⁺ 597.1968 (calcd. 597.1972). ¹H NMR (400 MHz,acetone-d₆) δ 8.12 (1H, s, H-2), 7.08 (1H, t, J=4.7 Hz, H-5), 6.98 (1H,ddd, J=8.0, 3.6, 1.3 Hz, H-5′), 6.93-6.75 (2H, m, H-2′, H-6′), 6.14-6.05(2H, n, H-9), 5.64-5.39 (2H, m, —OCH₂O—), 5.31 (1H, ddt, J=6.2, 4.6, 1.8Hz, H-1″), 5.13-5.02 (2H, m, OH), 4.52 (1H, d, J=6.0 Hz, H-3″), 3.96(3H, d, J=2.9 Hz, 3-OCH₃), 3.91-3.82 (1H, m, H-2″), 3.71 (3H, dd, J=5.5,3.1 Hz, 4-OCH₃), 3.55 (1H, dd, J=9.7, 5.9 Hz, H-4″), 3.37 (1H, ddd,J=9.6, 5.6, 3.8 Hz, H-5″), 2.38 (2H, td, J=7.4, 1.6 Hz, O-butyryl), 1.66(2H, dd, J=7.5, 1.6 Hz, O-butyryl), 1.35 (3H, dd, J=5.9, 1.8 Hz,O-butyryl), 0.97 (3H, td, J=7.4, 1.7 Hz, H-6″). ¹³C NMR (100 MHz,acetone-d₆) δ 173.6 (O-butyryl), 169.8 (C-9′), 152.9 (C-3), 151.4 (C-4),148.3 (C-3′), 148.3 (C-4′), 145.9 (C-7), 136.6 (C-7′), 131.4 (C-1),131.3 (C-6), 129.7 (C-1′), 128.2 (C-8), 124.6 (C-6′), 111.8 (C-8′),111.7 (C-2′), 108.6 (C-5′), 106.6 (C-2), 105.8 (C-1″), 102.4 (C-5),102.2 (—OCH₂O—), 78.3 (C-3″), 74.5 (C-2″), 73.7 (C-4″), 73.1 (C-5″),67.9 (C-9), 56.3 (3-OCH₃), 55.7 (4-OCH₃), 36.7 (O-butyryl), 19.0 (C-6″),18.1 (O-butyryl), 13.9 (O-butyryl).

3″-O-benzoyl-7-O-β-D-patentiflorin A (31d, yield 72% of two steps)

HR-EIMS: m/z [M+H]⁺ 631.1810 (calcd. 631.1816). ¹H NMR (400 MHz,acetone-d₆) δ 8.13 (1H, dt, J=6.2, 3.0 Hz, H-2), 8.11-8.04 (2H, m,O-Benzoyl), 8.01 (1H, dd, J=8.3, 2.7 Hz, O-Benzoyl), 7.51 (2H, d, J=6.1Hz, O-Benzoyl), 7.05 (I H, dq, J=10.2, 3.4 Hz, H-5), 6.95 (1H, dq,J=6.9, 3.8 Hz, H-5′), 6.90-6.73 (2H, m, H-2′, H-6′), 6.07 (2H, q, J=2.9,2.2 Hz, —OCH₂O—), 5.62-5.28 (4H, m, H-9, OH), 5.12 (1H, dd, J=7.8, 2.1Hz, H-1″), 4.07 (1H, q, J=9.1, 6.1 Hz, H-3″), 3.91 (3H, q, J=2.9, 2.5Hz, 3-OCH₃), 3.67 (3H, dt, J=7.1, 3.5 Hz, 4-OCH₃), 3.64-3.53 (3H, m,H-2″,4″ and 5″), 1.42-1.32 (3H, m, H-6″). ¹³C NMR (100 MHz, acetone-d₆)δ 169.9 (C-9′), 166.6 (O-benzoyl), 152.9 (C-3), 151.4 (C-4), 148.3(C-3′), 148.3 (C-4′), 146.0 (C-7), 136.6 (C-7′), 133.8 (O-benzoyl),133.7 (O-benzoyl), 131.6 (C-1), 131.5 (C-6), 131.3 (C-8), 130.5(O-benzoyl), 130.4 (O-benzoyl), 129.3 (O-benzoyl), 129.3 (O-benzoyl),128.2 (C-1′), 124.6 (C-6′), 111.8 (C-8′), 111.7 (C-2′), 108.7 (C-5′),106.6 (C-2), 105.8 (C-1″), 102.4 (C-5), 102.2 (—OCH₂O—), 79.5 (C-3″),74.5 (C-2″), 73.8 (C-4″), 73.1 (C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.7(4-OCH₃), 18.2 (C-6″).

3″-O-p-dimethylaminobenzoyl-7-O-β-D-patentiflorin A (31e, yield 51% oftwo steps)

HR-EIMS: m/z [M+H]⁺ 674.2099 (calcd. 674.2238). ¹H NMR (400 MHz,acetone-d₆) δ 8.21 (1H, d, J=1.7 Hz, H-2), 7.95-7.90 (2H, m,O-p-dimethylaminobenzoyl), 7.11 (1H, d, J=2.9 Hz, H-5), 7.01-6.96 (1H,m, H-5′), 6.93-6.81 (2H, m, H-2′, 6′), 6.79-6.75 (m, 2H,O-p-dimethylaminobenzoyl), 6.10 (2H, ddd, J=4.4, 1.7, 1.1 Hz, —OCH₂O—),5.60 (1H, d, J=15.0 Hz, H-1″), 5.50-5.42 (2H, m, H-9), 5.24 (1H, t,J=9.3 Hz, OH), 5.11 (1H, d, J=7.8 Hz, OH), 4.67 (1H, dd, J=5.7, 1.0 Hz,H-3″), 4.07-3.99 (1H, m, H-2″), 3.96 (3H, s, 3-OCH₃), 3.73 (3H, d, J=1.2Hz, 4-OCH₃), 3.65-3.57 (1H, m, H-5″), 3.52 (1H, td, J=9.2, 5.5 Hz,H-4″), 3.07 (6H, s, O-p-dimethylaminobenzoyl), 1.39 (3H, dd, J=6.0, 1.5Hz, H-6″). ¹³C NMR (100 MHz, acetone-d₆) δ 169.9 (C-9′), 167.4(O-p-dimethylaminobenzoyl), 154.6 (O-p-dimethylaminobenzoyl), 153.0(C-3), 151.5 (C-4), 148.4 (C-3′), 148.3 (C-4′), 146.1 (C-7), 136.6(C-7′), 132.3 (O-p-dimethylaminobenzoyl), 132.1(O-p-dimethylaminobenzoyl), 131.4 (C-1), 131.4 (C-6), 129.7 (C-8), 128.2(C-1′), 124.6 (C-6′), 120.0 (O-p-dimethylaminobenzoyl), 117.8(O-p-dimethylaminobenzoyl), 111.8 (C-8′), 111.5 (C-2′), 108.7 (C-5′),106.7 (C-2), 105.9 (C-1″), 102.5 (C-5), 102.2 (—OCH₂O—), 78.9 (C-3″),74.7 (C-2″), 74.0 (C-4″), 73.2 (C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.8(4-OCH₃), 40.1 (O-p-dimethylaminobenzoyl), 40.1(O-p-dimethylaminobenzoyl), 18.2 (C-6″).

3″-O-dimethylcarbamyl-7-O-β-D-patentiflorin A (31f, yield 66% of twosteps)

HR-EIMS: m/z [M+H]⁺ 674.2099 (calcd. 598.1925). ¹H NMR (400 MHz,acetone-d₆) δ 8.16 (1H, d, J=2.7 Hz, H-2), 7.07 (1H, d, J=4.9 Hz, H-5),6.97 (1H, dd, J=7.9, 2.7 Hz, H-5′), 6.92-6.78 (2H, m, H-2′, 6′),6.13-6.07 (2H, m, —OCH₂O—), 5.59-5.47 (2H, m, H-9), 5.42 (1H, dd,J=15.0, 0.8 Hz, H-1″), 5.02 (1H, d, J=7.8 Hz, H-3″), 4.80-4.72 (2H, m,OH), 3.95 (3H, s, 3-OCH₃), 3.90 (1H, ddd, J=9.4, 7.8, 4.5 Hz, H-2″),3.70 (3H, d, J=2.6 Hz, 4-OCH₃), 3.56-3.45 (1H, m, H-4″), 3.39 (1H, td,J=9.2, 4.4 Hz, H-5″), 3.02 (3H, s, O-dimethylcarbamyl), 2.92 (3H, s,O-dimethylcarbamyl), 1.35 (3H, dd, J=6.0, 1.5 Hz, H-6″). ¹³C NMR (100MHz, acetone-d₆) δ 169.9 (C-9′), 158.4 (O-dimethylcarbamyl), 152.9(C-3), 151.4 (C-4), 148.3 (C-3′), 148.2 (C-4′), 146.0 (C-7), 136.6(C-7′), 131.6 (C-1), 131.5 (C-6), 129.7 (C-8), 128.2 (C-1′), 124.6(C-6′), 111.8 (C-8′), 111.7 (C-2′), 108.6 (C-5′), 106.6 (C-2), 105.9(C-1″), 102.5 (C-5), 102.2 (—OCH₂O—), 81.4 (C-3″), 74.7 (C-2″), 74.0(C-4″), 73.0 (C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.7 (4-OCH₃), 36.7(O-dimethylcarbamyl), 36.2 (O-dimethylcarbamyl), 18.1 (C-6″).

3″-O-cyclopropanecarbonyl-7-O-4i-D-patentiflorin A (31g, yield 62% oftwo steps)

HR-EIMS: m/z [M+H]⁺ 595.1808 (calcd. 595.1816). ¹H NMR (400 MHz,acetone-d₆) δ 8.15 (1H, d, J=2.1 Hz, H-2), 7.09 (1H, d, J=4.2 Hz, H-5),6.98 (1H, dd, J=7.9, 2.8 Hz, H-5′), 6.91-6.79 (2H, m, H-2′, 6′),6.12-6.07 (2H, m, —OCH₂O—), 5.61-5.40 (3H, m, H-9, 1″), 5.10-5.01 (2H,m, OH), 4.69-4.61 (1H, m, H-3″), 3.95 (3H, s, 3-OCH₃), 3.89 (1H, ddd,J=12.2, 8.3, 4.1 Hz, H-2″), 3.71 (3H, d, J=2.3 Hz, 4-OCH₃), 3.59-3.49(1H, m, H-4″), 3.38 (1H, dd, J=10.7, 7.7 Hz, H-5″), 1.69 (1H, tt, J=7.9,4.7 Hz, O-cyclopropanecarbonyl), 1.35 (3H, dd, J=6.1, 1.5 Hz, H-6″),0.90-0.83 (4H, m, O-cyclopropanecarbonyl). ¹³C NMR (100 MHz, acetone-d₆)δ 175.1 (O-cyclopropanecarbonyl), 169.8 (C-9′), 152.9 (C-3), 151.4(C-4), 148.3 (C-3′), 148.3 (C-4′), 146.0 (C-7), 136.6 (C-7′), 131.4(C-1), 131.3 (C-6), 129.7 (C-8), 128.2 (C-1′), 124.6 (C-6′), 111.7(C-8′), 111.7 (C-2′), 108.6 (C-5′), 106.6 (C-2), 105.8 (C-1″), 102.5(C-5), 102.2 (—OCH₂O—), 78.7 (C-3″), 74.5 (C-2″), 73.7 (C-4″), 73.1(C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.7 (4-OCH₃), 18.1 (c-6″), 13.6(O-cyclopropanecarbonyl), 8.7 (O-cyclopropanecarbonyl), 8.2(O-cyclopropanecarbonyl).

3″-O-p-chlorobenzoyl-7-O-β-D-patentiflorin A (31h, yield 80% of twosteps)

HR-EIMS: m/z [M+H]⁺ 665.1413 (calcd. 665.1426). ¹H NMR (400 MHz,acetone-d₆) δ 8.15 (1H, d, J=2.4 Hz, H-2), 8.14-8.08 (2H, m,O-p-chlorobenzoyl), 7.61-7.55 (2H, m, O-p-chlorobenzoyl), 7.08 (1H, d,J=6.3 Hz, H-5), 7.00-6.95 (1H, m, H-5′), 6.93-6.78 (2H, m, H-2′, 6′),6.13-6.08 (2H, m, —OCH₂O—), 5.61-5.42 (3H, m, H-5, 1″), 5.35 (1H, t,J=9.2 Hz, OH), 5.15 (1H, dd, J=7.8, 1.2 Hz, OH), 4.80 (1H, dd, J=5.7,1.1 Hz, H-3″), 4.09 (1H, ddd, J=9.5, 7.8, 5.4 Hz, H-2″), 3.94 (3H, s,3-OCH₃), 3.70 (3H, d, J=3.7 Hz, 4-OCH₃), 3.68-3.54 (2H, m, H-4″, 5″),1.40 (3H, dd, J=5.9, 1.2 Hz, H-6″). ¹³C NMR (100 MHz, acetone-d₆) δ169.9 (C-9′), 165.8 (O-p-chlorobenzoyl), 152.9 (C-3), 151.4 (C-4), 148.3(C-3′), 148.3 (C-4′), 145.9 (C-7), 139.5 (O-p-chlorobenzoyl), 136.7(C-7′), 132.2 (O-p-chlorobenzoyl), 131.4 (C-1), 131.3 (C-6), 130.4(C-8), 129.5 (O-p-chlorobenzoyl), 128.2 (C-1′), 124.6 (C-6′), 111.8(C-8′), 111.7 (C-2′), 108.7 (C-5′), 106.6 (C-2), 105.7 (C-1″), 102.4(C-5), 102.2 (—OCH₂O—), 79.8 (C-3″), 74.4 (C-2″), 73.7 (C-4″), 73.1(C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.7 (4-OCH₃), 18.2 (C-6″).

3″-O-pyrazinecarbonyl-7-O-β-D-patentiflorin A (31i, yield 52% of twosteps)

HR-EIMS: m/z [M+H]⁺ 633.1709 (calcd. 633.1720). ¹H NMR (400 MHz,acetone-d₆) δ 8.89 (1H, d, J=1.5 Hz, O-pyrazinecarbonyl), 8.49 (1H, d,J=2.4 Hz, O-pyrazinecarbonyl), 8.43 (1H, dd, J=2.5, 1.5 Hz,O-pyrazinecarbonyl), 7.62 (I H, d, J=2.1 Hz, H-2), 6.60 (1H, dd, J=7.9,2.4 Hz, H-5′), 6.55 (1H, d, J=2.7 Hz, H-5), 6.49 (1H, dd, J=12.2, 1.7Hz, H-6′), 6.36 (1H, ddd, J=8.6, 7.9, 1.7 Hz, H-2′), 5.89 (1H, dd,J=5.6, 3.3 Hz, H-3″), 5.68 (2H, q, J=1.2 Hz, —OCH₂O—), 5.16-5.01 (3H, m,H-9, 1″), 4.76 (1H, t, J=9.3 Hz, OH), 4.60 (1H, dd, J=7.8, 4.9 Hz, OH),3.47 (3H, s, 3-OCH₃), 3.45-3.39 (1H, m, H-2″), 3.22 (3H, s, 4-OCH₃),3.16-3.07 (2H, m, H-4″, 5″), 0.86-0.80 (3H, m, H-6″). ¹³C NMR (100 MHz,acetone-d₆) δ 169.9 (C-9′), 168.6 (O-pyrazinecarbonyl), 153.0 (C-3),151.5 (C-4), 149.7 (C-3′), 148.3 (C-4′), 146.2 (O-pyrazinecarbonyl),145.9 (O-pyrazinecarbonyl), 145.5 (C-7), 144.8 (O-pyrazinecarbonyl),144.2 (O-pyrazinecarbonyl), 136.8 (C-7′), 131.5 (C-1), 131.4 (C-6),130.1 (C-8), 128.2 (C-1′), 124.6 (C-6′), 111.8 (C-8′), 111.7 (C-2′),108.7 (C-5′), 105.6 (C-1″), 102.4 (C-5), 102.2, (—OCH₂O—), 81.5 (C-3″),74.2 (C-2″), 73.5 (C-4″), 73.0 (C-5″), 67.9 (C-9), 56.3 (3-OCH₃), 55.8(4-OCH₃), 18.2 (C-6″).

3″-O-cyanoacetyl-7-O-β-D-patentiflorin A (31j, yield 49% of two steps)

HR-EIMS: m/z [M+H]) 594.1609 (calcd. 594.1612). ¹H NMR (400 MHz,acetone-d₆) δ 7.85 (1H, s, H-2), 7.09 (1H, m, H-5), 6.98 (1H, dd, J=7.9,2.5 Hz, H-5′), 6.89 (1H, dd, J=12.2, 1.6 Hz, H-2′), 6.85-6.79 (1H, m,H-6′), 6.15-6.02 (2H, m, —OCH₂O—), 5.70-5.37 (2H, m, H-9), 5.12-4.98(2H, m, H-1″, H-3″), 3.96 (3H, d, J=1.6 Hz, 3-OCH₃), 3.91 (1H, dd,J=9.5, 7.8 Hz, H-2″), 3.78-3.66 (3H, m, 4-OCH₃), 3.55 (1H, ddd, J=9.5,6.1, 1.2 Hz, H-4″), 3.35 (3H, m, O-cyanoacetyl, H-5″), 1.36 (3H, dd,J=6.0, 1.3 Hz, H-6″). ¹³C NMR (100 MHz, acetone-d₆) δ 169.9 (C-9′),167.8 (O-cyanoacetyl), 153.0 (C-3), 151.5 (C-4), 148.4 (C-3′), 148.3(C-4′), 146.0 (C-7), 136.6 (C-7′), 131.4 (C-1), 131.3 (C-6) 129.7 (C-8),128.2 (C-1′), 124.6 (C-6′), 113.8 (O-cyanoacetyl), 111.8 (C-2′), 111.8(C-8′), 108.7 (C-5′), 106.7 (C-2), 105.8 (C-1″), 102.5 (C-5), 102.2(—OCH₂O—), 79.2 (C-3″), 74.6 (C-2″), 73.9 (C-4″), 73.1 (C-5″), 67.9(C-9), 56.4 (3-OCH₃), 55.8 (4-OCH₃), 25.1 (O-cyanoacetyl), 18.2 (C-6″).

3″-O-trichloroacetyl-7-O-β-D-patentiflorin A (31k, yield 53% of twosteps)

HR-EIMS: m/z [M+H]⁺ 671.0325 (calcd. 671.0490). ¹H NMR (400 MHz,acetone-d₆) δ 8.15 (1H, d, J=1.5 Hz, H-2), 7.11 (1H, t, J=2.4 Hz, H-5),6.98 (1H, dd, J=7.9, 2.5 Hz, H-5′), 6.90 (1H, d, J=1.6 Hz, H-2′),6.88-6.80 (1H, m, H-6′), 6.10 (2H, dt, J=4.7, 1.2 Hz, —OCH₂O—),5.74-5.66 (1H, m, 4″-OH), 5.61-5.41 (2H, m, H-9), 5.12-5.02 (1H, m,2″-OH), 4.92 (1H, t, J=9.3 Hz, H-1″), 4.84 (1H, d, J=8.7 Hz, H-3″), 3.97(3H, d, J=1.3 Hz, 3-OCH₃), 3.91 (1H, ddd, J=9.5, 7.8, 5.4 Hz, H-2″),3.73 (3H, d, J=1.6 Hz, 4-OCH₃), 3.56 (1H, dq, J=12.2, 6.2 Hz, H-4″),3.42 (1H, td, J=9.3, 5.9 Hz, H-5″), 1.36 (3H, dd, J=6.1, 1.5 Hz, H-6″).¹³C NMR (100 MHz, acetone-d₆) δ 169.8 (C-9′), 162.5 (O-trichloroacetyl),153.0 (C-3), 151.5 (C-4), 148.4 (C-3′), 148.3 (C-4′), 145.9 (C-7), 136.7(C-7′), 131.5 (C-1), 131.4 (C-6), 129.7 (C-8), 128.2 (C-1′), 124.6(C-6′), 111.8 (C-8′), 111.7 (C-2′), 108.7 (C-5′), 106.7 (C-2), 105.7(C-1″), 102.5 (C-5), 102.2 (—OCH₂O—), 91.4 (O-trichloroacetyl), 79.6(C-3″), 74.1 (C-2″), 73.3 (C-4″), 73.1 (C-5″), 67.9 (C-9), 56.4(3-OCH₃), 55.8 (4-OCH₃), 18.1 (C-6″).

3″-O-benzyl-7-O-β-D-xylosyldiphyllin (311, yield 59% of two steps)

HR-EIMS: m/z [M+H]⁺ 603.1676 (calcd. 603.1866). ¹H NMR (400 MHz, CDCl₃)δ 7.85 (1H, s, H-2), 7.30 (5H, m, O-Bn), 7.01 (1H, m, H-5), 6.89 (1H, m,H-5′), 6.74 (1H, m, H-2′), 6.74 (1H, m, H-6′), 5.99 (2H, m, —OCH₂O—),5.38 (2H, m, H-9), 4.96 (1H, m, H-1″), 4.69 (2H, m, O-Bn), 3.96 (3H, s,3-OCH₃), 3.81 (1H, dd J=8.6, 7.2 Hz, H-3″), 3.73 (3H, s, 4-OCH₃), 3.60(3H, m, H-2″,4″,5″), 3.12 (1H, m, H-5″). ¹³C NMR (100 MHz, CDCl₁) δ169.9 (C-9′), 152.0 (C-3), 150.2 (C-4), 147.6 (C-3′), 147.5 (C-4′),144.3 (C-7), 137.6 (C-7′), 136.6 (O-Bn), 130.8 (C-1), 130.7 (C-6), 128.8(C-8), 128.8 (O-Bn), 128.5 (O-Bn), 128.3 (O-Bn), 128.3 (C-1′), 127.9(O-Bn), 127.0 (O-Bn), 123.6 (C-6′), 119.2 (C-8′), 110.7 (C-2′), 108.2(C-5′), 106.4 (C-2), 104.8 (C-1″), 101.3 (C-5), 100.9 (—OCH₂O—), 81.1(C-3″), 75.6 (C-2″), 75.3 (C-4″), 73.4 (C-5″), 73.04 (O-Bn), 67.4 (C-9),56.3 (3-OCH₃), 55.9 (4-OCH₃).

3″, 4″-Acetonide-4-O-β-D-glucopyranosyldiphyllin (32a)

To the solution of compound 26f (54.2 mg, 0.1 mmol) in 3 mL of2,2-dimethoxypropane (DMP) was added catalytic amount of TsOH (0.19 mg,0.001 mmol) (As illustrated in FIG. 6 ). After the reaction was stirredfor 12 h at r.t., the excess DMP was removed under evaporation inreduced pressure. The residue was purified by a silica gel columnseparation (n-hexane:ethyl acetate=1:1) to afford 32a as a white solid(11.4 mg, 20%): HR-EIMS: m/z [M+H]⁺ 583.1656 (calcd. 583.1816). ¹H NMR(400 MHz, Acetone-d₆) δ 8.22 (1H, s, H-2), 7.09 (1H, s, H-5), 6.97 (1H,dd, J=7.8, 2.2 Hz, H-5′), 6.87 (1H, dd, J=6.8, 2.2 Hz, H-2′), 6.81 (1H,q, J=7.4 Hz, H-6′), 6.10 (2H, d, J=3.6 Hz, —OCH₂O—), 5.52-5.34 (2H, m,H-9), 5.32 (1H, s, —OH), 5.03 (1H, dd, J=7.7, 2.8 Hz, H-1″), 4.66 (1H,s, —OH), 3.98 (3H, s, 3-OCH₃), 3.96-3.82 (2H, m, H-2″, H-3″), 3.79 (1H,td, J=7.9, 3.3 Hz, H-6″), 3.72 (3H, s, 4-OCH₃), 3.68-3.64 (2H, m, H-5″,H-6″), 3.34 (i H, q, J=7.9, 6.8 Hz, H-4″), 1.52 (3H, s,O-isopropylidene), 1.32 (3H, s, O-isopropylidene).

To the solution of compound 26g (50 mg, 0.09 mmol) in 3 mL of DMP wasadded catalytic amount of TsOH (0.19 mg, 0.001 mmol) (As illustrated inFIG. 6 ). After the reaction was stirred for 12 h at r.t., the excessDMP was removed under evaporation in reduced pressure. The residue waspurified by a silica gel column separation (45-52% ethyl acetate inn-hexane) to afford 32b (18 mg, 34%), 32c (20 mg, 38%) and 32d (2 mg,4%) with each as a white solid, respectively.

3″, 4″-Acetonide-4-O-β-D-galactopyranosyldiphyllin (32b)

HR-EIMS: m/z [M+H]⁺ 583.1666 (calcd. 583.1816). ¹H NMR (400 MHz,Acetone-d₆) δ 8.21 (1H, s, H-2), 7.09 (1H, s, H-5), 6.97 (1H, d, J=7.4Hz, H-5′), 6.90-6.75 (2H, m, H-2′, H-6′), 6.10 (2H, d, J=4.5 Hz,—OCH₂O—), 5.78 (1H, d, J=15.6 Hz, —OH), 5.48-5.39 (2H, m, H-9), 4.88(1H, d, J=9.5 Hz, H-1″), 4.28 (1H, d, J=5.9 Hz, H-3″), 4.23 (1H, t,J=6.0 Hz, H-4″), 4.07 (1H, q, J=5.7, 5.2 Hz, H-2″), 4.00 (3H, s,3-OCH₃), 3.96-3.83 (3H, m, H-5″, H-6″), 3.73 (3H, s, 4-OCH₃), 1.53 (3H,s, O-isopropylidene), 1.33 (3H, s, O-isopropylidene).

4″,6″-Acetonide-4-O-β-D-galactopyranosyldiphyllin (32c)

HR-EIMS: m/z [M+H]⁺ 583.1688 (calcd. 583.1816). ¹H NMR (400 MHz,Acetone-d₆) δ 8.28-8.19 (1H, m, H-2), 7.09 (1H, s, H-5), 6.98 (1H, dt,J=8.0, 1.1 Hz, H-5′), 6.90-6.78 (2H, m, H-2′, H-6′), 6.10 (2H, dt,J=5.0, 1.1 Hz, —OCH₂O—), 5.84-5.32 (2H, m, H-9), 4.94-4.83 (1H, m,H-1″), 4.32-4.20 (1H, m, H-6″), 4.04 (1H, dt, J=9.6, 7.6 Hz, H-2″), 4.00(3H, s, 3-OCH₃), 3.97-3.81 (3H, m, H-3″, H-4″, H-6″), 3.73 (3H, s,4-OCH₃), 3.66 (1H, tt, J=6.7, 4.3 Hz, H-5″), 1.53 (3H, s,O-isopropylidene), 1.33 (3H, s, O-isopropylidene).

2″,3″-Acetonide-4-O-β-D-galactopyranosyldiphyllin (32d)

HR-EIMS: m/z [M+H]⁺ 583.1648 (calcd. 583.1816). ¹H NMR (400 MHz,Acetone-d₆) δ 8.25 (1H, s, H-5), 7.11 (1H, s, H-5), 6.98 (1H, dd, J=7.8,1.0 Hz, H-5′), 6.90-6.86 (1H, m, H-2′), 6.83 (1H, dd, J=7.9, 1.7 Hz,H-6′), 6.10 (2H, dd, J=4.3, 1.0 Hz, —OCH₂O—), 5.65-5.45 (2H, m, H-9),4.96-4.90 (1H, m, H-1″), 4.26 (1H, d, J=3.5 Hz, H-3″), 4.15 (1H, dt,J=12.8, 2.1 Hz, H-2″), 4.01 (3H, s, 3-OCH₃), 3.93-3.83 (2H, m, H-6″),3.73 (3H, s, 4-OCH₃), 3.68 (1H, ddd, J=9.5, 7.9, 3.6 Hz, H-4″),3.57-3.50 (1H, m, H-5″), 1.47 (3H, s, O-isopropylidene), 1.41 (3H, s,O-isopropylidene).

Having now fully described the present invention in some detail by wayof illustration and examples for purposes of clarity of understanding,it will be obvious to one of ordinary skill in the art that the same canbe performed by modifying or changing the invention within a wide andequivalent range of conditions, formulations and other parameterswithout affecting the scope of the invention or any specific embodimentthereof, and that such modifications or changes are intended to beencompassed within the scope of the appended claims. The terms andexpressions which have been employed are used as terms of descriptionand not of limitation, and there is no intention that in the use of suchterms and expressions of excluding any equivalents of the features shownand described or portions thereof, but it is recognized that variousmodifications are possible within the scope of the invention claimed.Thus, it should be understood that although the present invention hasbeen specifically disclosed by preferred embodiments and optionalfeatures, modification and variation of the concepts herein disclosedmay be resorted to by those skilled in the art, and that suchmodifications and variations are considered to be within the scope ofthis invention.

As used herein, “comprising” is synonymous with “including.”“containing,” or “characterized by,” and is inclusive or open-ended anddoes not exclude additional, unrecited elements or method steps. As usedherein, “consisting of” excludes any element, step, or ingredient notspecified in the claim element. As used herein, “consisting essentiallyof” does not exclude materials or steps that do not materially affectthe basic and novel characteristics of the claim. In each instanceherein any of the terms “comprising”, “consisting essentially of” and“consisting of” may be replaced with either of the other two terms.

When a group of materials, compositions, components or compounds aredisclosed herein, it is understood that all individual members of thosegroups and all subgroups thereof are disclosed separately. When aMarkush group or other grouping is used herein, all individual membersof the group and all combinations and subcombinations possible of thegroup are intended to be individually included in the disclosure. Everyformulation or combination of components described or exemplified hereincan be used to practice the invention, unless otherwise stated. Whenevera range is given in the specification, for example, a temperature range,a time range, or a composition range, all intermediate ranges andsubranges, as well as all individual values included in the ranges givenare intended to be included in the disclosure. In the disclosure and theclaims, “and/or” means additionally or alternatively. Moreover, any useof a term in the singular also encompasses plural forms.

All references cited herein are hereby incorporated by reference intheir entirety to the extent that there is no inconsistency with thedisclosure of this specification. Some references provided herein areincorporated by reference to provide details concerning sources ofstarting materials, additional starting materials, additional reagents,additional methods of synthesis, additional methods of analysis,additional biological materials, additional cells, and additional usesof the invention. All headings used herein are for convenience only. Allpatents and publications mentioned in the specification are indicativeof the levels of skill of those skilled in the art to which theinvention pertains, and are herein incorporated by reference to the sameextent as if each individual publication, patent or patent applicationwas specifically and individually indicated to be incorporated byreference. References cited herein are incorporated by reference hereinin their entirety to indicate the state of the art as of theirpublication or filing date and it is intended that this information canbe employed herein, if needed, to exclude specific embodiments that arein the prior art. For example, when composition of matter is claimed, itshould be understood that compounds known and available in the art priorto applicants' invention, including compounds for which an enablingdisclosure is provided in the references cited herein, are not intendedto be included in the composition of matter claims herein.

What is claimed:
 1. A compound of Formula (I):

or a pharmaceutically acceptable salt or pro-drug thereof wherein, X isoxygen; R¹ is R¹⁵, R¹⁶; or —NH(CH₂R¹⁶); R², R⁵, R⁶, R¹⁰, R¹³, and R¹⁴are each hydrogen; R³ and R⁴ are each independently —O-alkyl; R⁷, R⁸,and R⁹ are each independently selected from the group consisting ofhydrogen and —O-alkyl; or R⁷ and R⁸ taken together with the carbon atomsto which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from C₁₋₆ alkyl; or R⁸ and R⁹ taken together with the carbonatoms to which they are attached to form a 5-6 membered heterocyclyloptionally substituted with 1, 2, 3, 4 or 5 group(s) independentlyselected from C₁₋₆ alkyl; R¹¹ and R¹² taken together form oxo; R¹⁵ is aglycosidic group represented by the Formula (V):

wherein, each of R¹⁹, R²¹, R²³ and R²⁵ is hydrogen; R²⁰ is selected fromthe group consisting of —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and—OC(O)OR²⁷; R²² is selected from the group consisting of —OR²⁷,—OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and —OC(O)OR²⁷; R²⁴ is selected from thegroup consisting of —OR²⁷, —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and —OC(O)OR²⁷;at least one of R²⁰, R²² and R²⁴ is selected from the group consistingof —OC(O)R²⁷, —OC(O)N(R²⁷)R²⁷, and —OC(O)OR²⁷; at least one of R²⁰, R²²and R²⁴ is hydroxyl; R²⁶ is hydrogen or methyl; R²⁷ for each occurrenceis independently selected from the group consisting of hydrogen,halogen, trichloromethyl, trifluoromethyl, cyano, nitro, —OR²⁹,—C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰,—N(R²⁹)S(O)₂R³⁰, hydrocarbyl optionally substituted with 1, 2, 3, 4 or 5group(s) independently selected from R²⁸, heterocyclcyl optionallysubstituted with 1, 2, 3, 4 or 5 group(s) independently selected fromR²⁸, and —(CH₂)_(k)-heterocyclyl optionally substituted with 1, 2, 3, 4or 5 group(s) independently selected from R²⁸, wherein k is an integerbetween 1-6; R²⁸ for each occurrence is independently selected fromhalogen, trichloromethyl, trifluoromethyl, cyano, nitro, oxo, ═NR²⁹,—OR²⁹, —C(O)R³⁰, —C(O)N(R²⁹)R³⁰, —C(O)OR²⁹, —OC(O)R²⁹, —S(O)₂R²⁹,—S(O)₂N(R²⁹)R³⁰, —N(R²⁹)R³⁰, —N(R²⁹)N(R²⁹)R³⁰, —N(R²⁹)C(O)R³⁰ and—N(R²⁹)S(O)₂R³⁰; and R²⁹ and R³⁰ for each occurrence are eachindependently hydrogen or selected from hydrocarbyl and heterocyclyl,either of which is optionally substituted with 1, 2, 3, 4 or 5 group(s)independently selected from halogen, cyano, amino, hydroxy, C₁₋₆ alkyland C₁₋₆ alkoxy; and R¹⁶ is selected from the group consisting of cyano,alkynyl, and alkynyl optionally substituted with a trialkylsilane. 2.The compound of claim 1, wherein R⁸ and R⁹ taken together with thecarbon atoms to which they are attached form a methylenedioxy ring. 3.The compound of claim 1, wherein R¹ is —C≡CH, —C≡C—Si(CH₃)₃, or—NH(CH₂—C≡CH).
 4. The compound of claim 3, wherein each of R³ and R⁴ is—OCH₃; and R⁸ and R⁹ taken together with the carbon atoms to which theyare attached form a methylenedioxy ring.
 5. The compound of claim 1,wherein the compound is selected from the group consisting of 19o, 21,and 23:


6. The compound of claim 1, wherein the glycosidic group is an isomerselected from the group consisting of α-D, α-L, β-D, and β-L.
 7. Thecompound of claim 1, wherein the glycosidic group is a monosaccharideselected from the group consisting of an α-L isomer and an β-L isomer.8. The compound of claim 1, wherein the compound is selected from thegroup consisting of 27aa, 27ab, 27ac, 27ad, 27ae, 27af, 27ba, 27bb,27bc, 27bd, 27be, 27bf, 28ab1, 28ab2, 28bb1, 28bb2, 29a, 29b, 31a, 31b,31c, 31d, 31e, 31f, 31g, 31h, 31i, 31j, and 31k:


9. A pharmaceutical composition comprising a compound of claim 1 atleast one pharmaceutically acceptable excipient.
 10. The compound ofclaim 8, wherein the compound is selected from the group consisting of27ba, 27bb, 27bc, 27bd, 27be, 27bf, 28ab2, 29b, 31b, 31c, 31d, 31e, 31i,and 31j.
 11. A method of treating an HIV infection in a subject in needthereof, the method comprising administering a therapeutically effectiveamount of a compound of claim 1 to the subject.